(+)-2-[1-(3-Ethoxy-4methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3,-dione:methods of using and compositions thereof

ABSTRACT

Stereomerically pure (+)-2-[1-(3-Ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dione, substantially free of its (−) isomer, and prodrugs, metabolites, polymorphs, salts, solvates, hydrates, and clathrates thereof are discussed. Also discussed are methods of using and pharmaceutical compositions comprising the (+) enantiomer of 2-[1-(3-Ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dione are disclosed. The methods include methods of treating and/or preventing disorders ameliorated by the reduction of levels of TNF-α or the inhibition of PDE4.

This application claims the benefit of U.S. Provisional Application No.60/366,515 filed Mar. 20, 2002 and U.S. Provisional Application No.60/438,450 filed Jan. 7, 2003 both of which are incorporated herein byreference in their entireties.

1. FIELD OF INVENTION

The invention relates to methods of using and compositions comprisingthe (+) enantiomer of2-[1-(3-Ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dione.

2. BACKGROUND OF THE INVENTION

Tumor necrosis factor alpha, (TNF-α) is a cytokine that is releasedprimarily by mononuclear phagocytes in response to immunostimulators.TNF-α is capable of enhancing most cellular processes, such asdifferentiation, recruitment, proliferation, and proteolyticdegradation. At low levels, TNF-α confers protection against infectiveagents, tumors, and tissue damage. But TNF-α also has a role in manydiseases. When administered to mammals or humans, TNF-α causes oraggravates inflammation, fever, cardiovascular effects, hemorrhage,coagulation, and acute phase responses similar to those seen duringacute infections and shock states. Enhanced or unregulated TNF-αproduction has been implicated in a number of diseases and medicalconditions, for example, cancers, such as solid tumors and blood-borntumors; heart disease, such as congestive heart failure; and viral,genetic, inflammatory, allergic, and autoimmune diseases.

Adenosine 3′,5′-cyclic monophosphate (cAMP) also plays a role in manydiseases and conditions, such as but not limited to asthma andinflammation, and other conditions (Lowe and Cheng, Drugs of the Future,17(9), 799-807, 1992). It has been shown that the elevation of cAMP ininflammatory leukocytes inhibits their activation and the subsequentrelease of inflammatory mediators, including TNF-α and NF-κB. Increasedlevels of cAMP also leads to the relaxation of airway smooth muscle.

It is believed that the primary cellular mechanism for the inactivationof cAMP is the breakdown of cAMP by a family of isoenzymes referred toas cyclic nucleotide phosphodiesterases (PDE) (Beavo and Reitsnyder,Trends in Pharm., 11, 150-155, 1990). There are eleven known PDEfamilies. It is recognized, for example, that the inhibition of PDE typeIV is particularly effective in both the inhibition of inflammatorymediator release and the relaxation of airway smooth muscle (Verghese,et al., Journal of Pharmacology and Experimental Therapeutics, 272(3),1313-1320, 1995). Thus, compounds that inhibit PDE4 (PDE IV)specifically, may inhibit inflammation and aid the relaxation of airwaysmooth muscle with a minimum of unwanted side effects, such ascardiovascular or anti-platelet effects. Currently used PDE4 inhibitorslack the selective action at acceptable therapeutic doses.

Cancer is a particularly devastating disease, and increases in bloodTNF-α levels are implicated in the risk of and the spreading of cancer.Normally, in healthy subjects, cancer cells fail to survive in thecirculatory system, one of the reasons being that the lining of bloodvessels acts as a barrier to tumor-cell extravasation. But increasedlevels of cytokines have been shown to substantially increase theadhesion of cancer cells to endothelium in vitro. One explanation isthat cytokines, such as TNF-α, stimulate the biosynthesis and expressionof a cell surface receptors called ELAM-1 (endothelial leukocyteadhesion molecule). ELAM-1 is a member of a family of calcium-dependentcell adhesion receptors, known as LEC-CAMs, which includes LECAM-1 andGMP-140. During an inflammatory response, ELAM-1 on endothelial cellsfunctions as a “homing receptor” for leukocytes. Recently, ELAM-1 onendothelial cells was shown to mediate the increased adhesion of coloncancer cells to endothelium treated with cytokines (Rice et al., 1989,Science 246:1303-1306).

Inflammatory diseases such as arthritis, related arthritic conditions(e.g., osteoarthritis and rheumatoid arthritis), inflammatory boweldisease (e.g., Crohn's disease and ulcerative colitis), sepsis,psoriasis, atopic dermatitis, contact dermatitis, and chronicobstructive pulmonary disease, chronic inflammatory pulmonary diseasesare also prevalent and problematic ailments. TNF-α plays a central rolein the inflammatory response and the administration of their antagonistsblock chronic and acute responses in animal models of inflammatorydisease.

Enhanced or unregulated TNF-α production has been implicated in viral,genetic, inflammatory, allergic, and autoimmune diseases. Examples ofsuch diseases include but are not limited to: HIV; hepatitis; adultrespiratory distress syndrome; bone-resorption diseases; chronicobstructive pulmonary diseases; chronic pulmonary inflammatory diseases;asthma, dermatitis; cystic fibrosis; septic shock; sepsis; endotoxicshock; hemodynamic shock; sepsis syndrome; post ischemic reperfusioninjury; meningitis; psoriasis; fibrotic disease; cachexia; graftrejection; auto-immune disease; rheumatoid spondylitis; arthriticconditions, such as rheumatoid arthritis and osteoarthritis;osteoporosis; Crohn's disease; ulcerative colitis; inflammatory-boweldisease; multiple sclerosis; systemic lupus erythrematosus; ENL inleprosy; radiation damage; asthma; and hyperoxic alveolar injury. Traceyet al., 1987, Nature 330:662-664 and Hinshaw et al., 1990, Circ. Shock30:279-292 (endotoxic shock); Dezube et al., 1990, Lancet, 335:662(cachexia); Millar et al., 1989, Lancet 2:712-714 and Ferrai-Balivieraet al., 1989, Arch. Surg. 124:1400-1405 (adult respiratory distresssyndrome); Bertolini et al., 1986, Nature 319:516-518, Johnson et al.,1989, Endocrinology 124:1424-1427, Holler et al., 1990, Blood75:1011-1016, and Grau et al., 1989, N. Engl. J. Med. 320:1586-1591(bone resorption diseases); Pignet et al., 1990, Nature, 344:245-247,Bissonnette et al., 1989, Inflammation 13:329-339 and Baughman et al.,1990, J. Lab. Clin. Med. 115:36-42 (chronic pulmonary inflammatorydiseases); Elliot et al., 1995, Int. J. Pharmac. 17:141-145 (rheumatoidarthritis); von Dullemen et al., 1995, Gastroenterology, 109:129-135(Crohn's disease); Duh et al., 1989, Proc. Nat. Acad. Sci. 86:5974-5978,Poll et al., 1990, Proc. Nat. Acad. Sci. 87:782-785, Monto et al., 1990,Blood 79:2670, Clouse et al., 1989, J. Immunol. 142, 431-438, Poll etal., 1992, AIDS Res. Hum. Retrovirus, 191-197, Poli et al. 1990, Proc.Natl. Acad. Sci. 87:782-784, Folks et al., 1989, PNAS 86:2365-2368 (HIVand opportunistic infections resulting from HIV).

Pharmaceutical compounds that can block the activity or inhibit theproduction of certain cytokines, including TNF-α, may be beneficialtherapeutics. Many small-molecule inhibitors have demonstrated anability to treat or prevent inflammatory diseases implicated by TNF-α(for a review, see Lowe, 1998 Exp. Opin. Ther. Patents 8:1309-1332). Onesuch class of molecules are the substituted phenethylsulfones describedin U.S. Pat. No. 6,020,358.

3. SUMMARY OF THE INVENTION

This invention relates to methods of treating diseases and disordersutilizing an enantiomer of a substituted phenethylsulfone compound andpharmaceutically acceptable salts, hydrates, solvates, clathrates,prodrugs and polymorphs thereof and methods for reducing the level ofcytokines and their precursors in mammals. The invention also relates topharmaceutical compositions comprising an enantiomer of2-[1-(3-Ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dioneand a pharmaceutically acceptable carrier. The invention further relatesto an enantiomer of2-[1-(3-Ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dionesubstantially free of its other enantiomer.

This invention particularly relates to the (+) enantiomer of2-[1-(3-Ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dione.This compound is believed to have increased potency and other benefitsas compared to itsracemate—2-[1-(3-Ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dione.

The invention encompasses the use of the (+) enantiomer of2-[1-(3-Ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dionefor treating or preventing diseases or disorders ameliorated by theinhibition of TNF-α production in mammals. In certain embodiments, thistreatment includes the reduction or avoidance of adverse effects. Suchdisorders include, but are not limited to, cancers, including, but notlimited to cancer of the head, thyroid, neck, eye, skin, mouth, throat,esophagus, chest, bone, blood, bone marrow, lung, colon, sigmoid,rectum, stomach, prostate, breast, ovaries, kidney, liver, pancreas,brain, intestine, heart, adrenal, subcutaneous tissue, lymph nodes,heart, and combinations thereof. Specific cancers that can be treated bythis method are multiple myeloma, malignant melanoma, malignant glioma,leukemia and solid tumors.

The invention also encompasses the use of the (+) enantiomer of2-[1-(3-Ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dionein the treatment or prevention of heart disease, including, but notlimited to congestive heart failure, cardiomyopathy, pulmonary edema,endotoxin-mediated septic shock, acute viral myocarditis, cardiacallograft rejection, and myocardial infarction.

The invention also encompasses the use of the (+) enantiomer of2-[1-(3-Ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dioneto treat diseases or disorders ameliorated by the inhibition of PDE4.For example, the compounds and compositions of the invention may beuseful to treat or prevent viral, genetic, inflammatory, allergic, andautoimmune diseases. Examples of such diseases include, but are notlimited to: HIV; hepatitis; adult respiratory distress syndrome;bone-resorption diseases; chronic obstructive pulmonary diseases;chronic pulmonary inflammatory diseases; dermatitis; inflammatory skindisease, atopic dermatitis, cystic fibrosis; septic shock; sepsis;endotoxic shock; hemodynamic shock; sepsis syndrome; post ischemicreperfusion injury; meningitis; psoriasis; fibrotic disease; cachexia;graft rejection including graft versus host disease; auto-immunedisease; rheumatoid spondylitis; arthritic conditions, such asrheumatoid arthritis and osteoarthritis; osteoporosis; Crohn's disease;ulcerative colitis; inflammatory-bowel disease; multiple sclerosis;systemic lupus erythrematosus; erythema nodosum leprosum (ENL) inleprosy; radiation damage; asthma; and hyperoxic alveolar injury.

In yet another embodiment, the stereomerically pure (+) enantiomer of2-[1-(3-Ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dioneis also useful in the treatment or prevention of microbial infections orthe symptoms of microbial infections including, but not limited to,bacterial infections, fungal infections, malaria, mycobacterialinfection, and opportunistic infections resulting from HIV.

The invention further encompasses pharmaceutical compositions and singleunit dosage forms comprising an enantiomer of2-[1-(3-Ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dioneand pharmaceutically acceptable polymorphs, prodrugs, salts, hydrates,clathrates, and solvates thereof.

In a separate embodiment, the invention encompasses the (+) enantiomerof2-[1-(3-Ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dione.

In a further embodiment, the invention encompasses a method of producinga stereomerically pure enantiomer of2-[1-(3-Ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dionewhich comprises contacting1-(3-Ethoxy-4-methoxy-phenyl)-2-methanesulfonyl-ethylamine with a chiralamino acid and contacting the product of the first step withN-(1,3-Dioxo-1,3-dihydro-isobenzofuran-4-yl)-acetamide. In a relatedembodiment the invention encompasses a chiral salt of1-(3-Ethoxy-4-methoxy-phenyl)-2-methanesulfonyl-ethylamine.

3.1. BRIEF DESCRIPTION OF THE FIGURES

FIG. 1. illustrates the preparation of the (+) enantiomer of2-[1-(3-Ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dione.

FIG. 2. illustrates the effect of the enantiomer of the invention onLPS-induced neutrophilia in the lungs of conscious ferrets.

3.2. DEFINITIONS

As used herein, term “Compound A” refers to an enantiomerically pureform of2-[1-(3-ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dionewhich comes off of an HPLC column at about 25.4 minutes when that columnis a 150 mm×4.6 mm Ultron Chiral ES-OVS chiral HPLC column (AgilentTechnology), the eluent is 15:85 ethanol: 20 mM KH₂PO₄ at pH 3.5, andthe observation wavelength is 240 nm. The ¹H NMR spectrum of compound Ais substantially as follows: δ(CDCl₃): 1.47 (t, 3H), 2.26 (s, 3H), 2.87(s, 3H), 3.68-3.75 (dd, 1H), 3.85 (s, 3H), 4.07-4.15 (q, 2H), 4.51-4.61(dd, 1H), 5.84-5.90 (dd, 1H), 6.82-8.77 (m, 6H), 9.46 (s, 1H). The ¹³CNMR spectrum of Compound A is substantially as follows δ(DMSO-d₆):14.66, 24.92, 41.61, 48.53, 54.46, 55.91, 64.51, 111.44, 112.40, 115.10,118.20, 120.28, 124.94, 129.22, 131.02, 136.09, 137.60, 148.62, 149.74,167.46, 169.14, 169.48. Compound A dissolved in methanol also rotatesplane polarized light in the (+) direction.

Without being limited by theory, Compound A is believed to beS-{2-[1-(3-ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dione},which has the following structure:

As used herein, the term “patient” refers to a mammal, particularly ahuman.

As used herein, the term “pharmaceutically acceptable salts” refer tosalts prepared from pharmaceutically acceptable non-toxic acids or basesincluding inorganic acids and bases and organic acids and bases.Suitable pharmaceutically acceptable base addition salts for thecompound of the present invention include metallic salts made fromaluminum, calcium, lithium, magnesium, potassium, sodium and zinc ororganic salts made from lysine, N,N′-dibenzylethylenediamine,chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine(N-methylglucamine) and procaine. Suitable non-toxic acids include, butare not limited to, inorganic and organic acids such as acetic, alginic,anthranilic, benzenesulfonic, benzoic, camphorsulfonic, citric,ethenesulfonic, formic, fumaric, furoic, galacturonic, gluconic,glucuronic, glutamic, glycolic, hydrobromic, hydrochloric, isethionic,lactic, maleic, malic, mandelic, methanesulfonic, mucic, nitric, pamoic,pantothenic, phenylacetic, phosphoric, propionic, salicylic, stearic,succinic, sulfanilic, sulfuric, tartaric acid, and p-toluenesulfonicacid. Specific non-toxic acids include hydrochloric, hydrobromic,phosphoric, sulfuric, and methanesulfonic acids. Examples of specificsalts thus include hydrochloride and mesylate salts.

As used herein and unless otherwise indicated, the term “prodrug” meansa derivative of a compound that can hydrolyze, oxidize, or otherwisereact under biological conditions (in vitro or in vivo) to provide thecompound. Examples of prodrugs include, but are not limited to,derivatives and metabolites of Compound A that include biohydrolyzablemoieties such as biohydrolyzable amides, biohydrolyzable esters,biohydrolyzable carbamates, biohydrolyzable carbonates, biohydrolyzableureides, and biohydrolyzable phosphate analogues. Prodrugs can typicallybe prepared using well-known methods, such as those described by 1Burger's Medicinal Chemistry and Drug Discovery, 172-178, 949-982(Manfred E. Wolff ed., 5th ed. 1995).

As used herein and unless otherwise indicated, the terms“biohydrolyzable amide,” “biohydrolyzable ester,” “biohydrolyzablecarbamate,” “biohydrolyzable carbonate,” “biohydrolyzable ureide,”“biohydrolyzable phosphate” mean an amide, ester, carbamate, carbonate,ureide, or phosphate, respectively, of a compound that either: 1) doesnot interfere with the biological activity of the compound but canconfer upon that compound advantageous properties in vivo, such asuptake, duration of action, or onset of action; or 2) is biologicallyinactive but is converted in vivo to the biologically active compound.Examples of biohydrolyzable esters include, but are not limited to,lower alkyl esters, alkoxyacyloxy esters, alkyl acylamino alkyl esters,and choline esters. Examples of biohydrolyzable amides include, but arenot limited to, lower alkyl amides, α-amino acid amides, alkoxyacylamides, and alkylaminoalkylcarbonyl amides. Examples of biohydrolyzablecarbamates include, but are not limited to, lower alkylamines,substituted ethylenediamines, aminoacids, hydroxyalkylamines,heterocyclic and heteroaromatic amines, and polyether amines.

As used herein and unless otherwise indicated, the term “stereomericallypure” means a composition that comprises one stereoisomer of a compoundand is substantially free of other stereoisomers of that compound. Forexample, a stereomerically pure composition of a compound having onechiral center will be substantially free of the opposite enantiomer ofthe compound. A stereomerically pure composition of a compound havingtwo chiral centers will be substantially free of other diastereomers ofthe compound. A typical stereomerically pure compound comprises greaterthan about 80% by weight of one stereoisomer of the compound and lessthan about 20% by weight of other stereoisomers of the compound, morepreferably greater than about 90% by weight of one stereoisomer of thecompound and less than about 10% by weight of the other stereoisomers ofthe compound, even more preferably greater than about 95% by weight ofone stereoisomer of the compound and less than about 5% by weight of theother stereoisomers of the compound, and most preferably greater thanabout 97% by weight of one stereoisomer of the compound and less thanabout 3% by weight of the other stereoisomers of the compound.

As used herein and unless otherwise indicated, the term“enantiomerically pure” means a stereomerically pure composition of acompound having one chiral center.

As used herein, term “adverse effects” includes, but is not limited togastrointestinal, renal and hepatic toxicities, leukopenia, increases inbleeding times due to, e.g., thrombocytopenia, and prolongation ofgestation, nausea, vomiting, somnolence, asthenia, dizziness,teratogenicity, extra-pyramidal symptoms, akathisia, cardiotoxicityincluding cardiovascular disturbances, inflammation, male sexualdysfunction, and elevated serum liver enzyme levels. The term“gastrointestinal toxicities” includes but is not limited to gastric andintestinal ulcerations and erosions. The term “renal toxicities”includes but is not limited to such conditions as papillary necrosis andchronic interstitial nephritis.

As used herein and unless otherwise indicated, the phrases “reduce oravoid adverse effects” and “reducing or avoiding adverse effects” meanthe reduction of the severity of one or more adverse effects as definedherein.

It should be noted that if there is a discrepancy between a depictedstructure and a name given that structure, the depicted structure is tobe accorded more weight. In addition, if the stereochemistry of astructure or a portion of a structure is not indicated with, forexample, bold or dashed lines, the structure or portion of the structureis to be interpreted as encompassing all stereoisomers of it.

4. DETAILED DESCRIPTION OF THE INVENTION

This invention relates to stereomerically pure Compound A, which is anenantiomer of2-[1-(3-Ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dione,substantially free of its other enantiomer, as well as novel methodsusing, and compositions comprising stereomerically pure Compound A. Forexample, the present invention encompasses the in vitro and in vivo useof Compound A, and the incorporation of Compound A into pharmaceuticalcompositions and single unit dosage forms useful in the treatment andprevention of a variety of diseases and disorders. Diseases anddisorders which are ameliorated by the reduction of levels of TNF-α orinhibition of PDE4 are well known in the art and are described herein.Specific methods of the invention reduce or avoid the adverse effectsassociated with compounds used as TNF-α inhibitor. Other specificmethods of the invention reduce or avoid the adverse effects associatedwith use of racemic2-[1-(3-Ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dione.

Specific methods of the invention include methods of treating orpreventing diseases and disorders including, but not limited to, solidtumor cancers, blood-born cancers and inflammatory diseases.

Pharmaceutical and dosage forms of the invention, which compriseCompound A or a pharmaceutically acceptable polymorph, prodrug, salt,clathrate, solvate or hydrate thereof, can be used in the methods of theinvention.

Without being limited by theory, it is believed that Compound A caninhibit TNF-α production. Consequently, a first embodiment of theinvention relates to a method of inhibiting TNF-α production whichcomprises contacting a cell exhibiting abnormal TNF-α production with aneffective amount of stereomerically pure Compound A, or apharmaceutically acceptable prodrug, metabolite, polymorph, salt,solvate, hydrate, or clathrate thereof. In a particular embodiment, theinvention relates to a method of inhibiting TNF-α production whichcomprises contacting a mammalian cell exhibiting abnormal TNF-αproduction with an effective amount of stereomerically pure Compound A,or a pharmaceutically acceptable prodrug, metabolite, polymorph, salt,solvate, hydrate, or clathrate thereof.

The invention also relates to a method of treating or preventingdisorders ameliorated by the reduction of levels of TNF-α in a patientwhich comprises administering to a patient in need of such treatment orprevention a therapeutically or prophylactically effective amount ofstereomerically pure compound A, or a pharmaceutically acceptableprodrug, metabolite, polymorph, salt, solvate, hydrate, or clathratethereof.

A further embodiment of the invention relates to a method of treating orpreventing cancer, including but not limited to, solid tumor, blood-borntumor, leukemias, and in particular, multiple myeloma in a patient whichcomprises administering to a patient in need of such treatment orprevention a therapeutically effective amount of stereomerically purecompound A, or a pharmaceutically acceptable prodrug, metabolite,polymorph, salt, solvate, hydrate, or clathrate thereof; in particularwherein the patient is a mammal.

In another embodiment, the invention relates to a method of inhibitingPDE4 which comprises contacting PDE4 with an effective amount ofstereomerically pure Compound A, or a pharmaceutically acceptableprodrug, metabolite, polymorph, salt, solvate, hydrate, or clathratethereof.

In another embodiment, the invention relates to a method of controllingcAMP levels in a cell which comprises contacting a cell with aneffective amount of stereomerically pure Compound A, or apharmaceutically acceptable prodrug, metabolite, polymorph, salt,solvate, hydrate, or clathrate thereof. As used herein the term“controlling cAMP levels” includes preventing or reducing the rate ofthe breakdown of Adenosine 3′,5′-cyclic monophosphate (cAMP) in a cellor increasing the amount of Adenosine 3′,5′-cyclic monophosphate presentin a cell, preferably a mammalian cell, more preferably a human cell. Ina particular method, the rate of cAMP breakdown is reduced by about 10,25, 50, 100, 200, or 500 percent as compared to the rate in comparablecells which have not been contacted with a compound of the invention.

A further embodiment of the invention relates to a method of treating orpreventing diseases or disorders ameliorated by the inhibition of PDE4in a patient which comprises administering to a patient in need of suchtreatment or prevention a therapeutically or prophylactically effectiveamount of stereomerically pure Compound A, or a pharmaceuticallyacceptable prodrug, metabolite, polymorph, salt, solvate, hydrate, orclathrate thereof. Disorders ameliorated by the inhibition of PDE4include, but are not limited to, asthma, inflammation (e.g.,inflammation due to reperfusion), chronic or acute obstructive pulmonarydiseases, chronic or acute pulmonary inflammatory diseases, inflammatorybowel disease, Crohn's Disease, Bechet's Disease, or colitis.

A further embodiment of the invention relates to a method of treating orpreventing depression, asthma, inflammation (e.g., contact dermatitis,atopic dermatitis, psoriasis, rheumatoid arthritis, osteoarthritis,inflammatory skin disease, inflammation due to reperfusion), chronic oracute obstructive pulmonary diseases, chronic or pulmonary inflammatorydiseases, inflammatory bowel disease, Crohn's Disease, Bechet's Diseaseor colitis in a patient which comprises administering to a patient inneed of such treatment or prevention a therapeutically orprophylactically effective amount of stereomerically pure Compound A, ora pharmaceutically acceptable prodrug, metabolite, polymorph, salt,solvate, hydrate, or clathrate thereof; in particular wherein thepatient is a mammal.

A separate embodiment of the invention encompasses methods of treatingor preventing Myelodysplastic syndrome (MDS) which comprisesadministering to a patient in need of such treatment or prevention atherapeutically or prophylactically effective amount of stereomericallypure Compound A, or a pharmaceutically acceptable salt, solvate,hydrate, stereoisomer, clathrate, or prodrug thereof. MDS refers to adiverse group of hematopoietic stem cell disorders. MDS is characterizedby a cellular marrow with impaired morphology and maturation(dysmyelopoiesis), peripheral blood cytopenias, and a variable risk ofprogression to acute leukemia, resulting from ineffective blood cellproduction. See The Merck Manual 953 (17th ed. 1999) and List et al.,1990, J. Clin. Oncol. 8:1424.MDS

A separate embodiment of the invention encompasses methods of treatingor preventing Myeloproliferative disease (MPD) which comprisesadministering to a patient in need of such treatment or prevention atherapeutically or prophylactically effective amount of stereomericallypure Compound A, or a pharmaceutically acceptable salt, solvate,hydrate, stereoisomer, clathrate, or prodrug thereof. Myeloproliferativedisease (MPD) refers to a group of disorders characterized by clonalabnormalities of the hematopoietic stem cell. See e.g., Current MedicalDiagnosis & Treatment, pp. 499 (37th ed., Tierney et al. ed, Appleton &Lange, 1998).

The invention also encompasses a method of treating, preventing ormanaging complex regional pain syndrome, which comprises administeringto a patient in need of such treatment, prevention or management atherapeutically or prophylactically effective amount of astereomerically pure Compound A, or a pharmaceutically acceptable salt,solvate, hydrate, stereoisomer, clathrate, or prodrug thereof. In aspecific embodiment, the administration is before, during or aftersurgery or physical therapy directed at reducing or avoiding a symptomof complex regional pain syndrome in the patient.

In particular methods of the invention, stereomerically pure Compound A,or a pharmaceutically acceptable polymorph, prodrug, salt, solvate,hydrate, or clathrate thereof, is adjunctively administered with atleast one additional therapeutic agent. Examples of additionaltherapeutic agents include, but are not limited to, anti-cancer drugs,anti-inflammatories, antihistamines and decongestants.

4.1. Synthesis And Preparation

Racemic2-[1-(3-Ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dioneis readily prepared using the methods in U.S. Pat. No. 6,020,358, whichis incorporated herein by reference.

Compound A can be isolated from the racemic compound by techniques knownin the art. Examples include, but are not limited to, the formation ofchiral salts and the use of chiral or high performance liquidchromatography “HPLC” and the formation and crystallization of chiralsalts. See, e.g., Jacques, J., et al., Enantiomers, Racemates andResolutions(Wiley-Interscience, New York, 1981); Wilen, S. H., et al.,Tetrahedron 33:2725 (1977); Eliel, E. L., Stereochemistry of CarbonCompounds (McGraw-Hill, NY, 1962); and Wilen, S. H., Tables of ResolvingAgents and Optical Resolutions p. 268 (E. L. Eliel, Ed., Univ. of NotreDame Press, Notre Dame, Ind., 1972).

In a specific method, Compound A is synthesized from 3-acetamidophthalicanhydride and a chiral amino acid salt of(S)-2-(3-ethoxy-4-methoxyphenyl)-1-(methylsulphonyl)-eth-2-ylamine.Chiral amino acid salts of(S)-2-(3-ethoxy-4-methoxyphenyl)-1-(methylsulphonyl)-eth-2-ylamineinclude, but not limited to salts formed with the L isomers of alanine,arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid,glycine, histidine, isoleucine, leucine, lysine, methionine,phenylalanine, proline, serine, threonine, tryptophan, tyrosine, valine,ornithine, 4-aminobutyric acid, 2 amino isobutyric acid, 3 aminopropionic acid, ornithine, norleucine, norvaline, hydroxyproline,sarcosine, citrulline, cysteic acid, t-butylglycine, t-butylalanine,phenylglycine, cyclohexylalanine, and N-acetyl-leucine. A specificchiral amino acid salt is(S)-2-(3-ethoxy-4-methoxyphenyl)-1-(methylsulphonyl)-eth-2-ylamineN-acetyl-L-leucine salt, which is resolved from2-(3-ethoxy-4-methoxyphenyl)-1-(methylsulphonyl)-eth-2-ylamine andN-acetyl-L-leucine in methanol.

4.2. Methods Of Treatment

The invention encompasses methods of treating and preventing diseases ordisorders ameliorated by the reduction of levels of TNF-α in a patientwhich comprise administering to a patient in need of such treatment orprevention a therapeutically effective amount of stereomerically pureCompound A, or a pharmaceutically acceptable prodrug, metabolite,polymorph, salt, solvate, hydrate, or clathrate thereof.

Disorders ameliorated by the inhibition of TNF-α include, but are notlimited to: heart disease, such as congestive heart failure,cardiomyopathy, pulmonary edema, endotoxin-mediated septic shock, acuteviral myocarditis, cardiac allograft rejection, and myocardialinfarction; solid tumors, including but not limited to, sarcoma,carcinomas, fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma,osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma,lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma,Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon carcinoma,pancreatic cancer, breast cancer, ovarian cancer, prostate cancer,squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweatgland carcinoma, sebaceous gland carcinoma, papillary carcinoma,papillary adenocarcinomas, cystadenocarcinoma, medullary carcinoma,bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile ductcarcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilms' tumor,cervical cancer, testicular tumor, lung carcinoma, small cell lungcarcinoma, bladder carcinoma, epithelial carcinoma, glioma, astrocytoma,medulloblastoma, craniopharyngioma, ependymoma, Kaposi's sarcoma,pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma,menangioma, melanoma, neuroblastoma, and retinoblastoma; and blood-borntumors including but not limited to, acute lymphoblastic leukemia “ALL”,acute lymphoblastic B-cell leukemia, acute lymphoblastic T-cellleukemia, acute myeloblastic leukemia “AML”, acute promyelocyticleukemia “APL”, acute monoblastic leukemia, acute erythroleukemialeukemia, acute megakaryoblastic leukemia, acute myelomonocyticleukemia, acute nonlymphocytic leukemia, acute undifferentiatedleukemia, chronic myelocytic leukemia “CML”, chronic lymphocyticleukemia “CLL”, hairy cell leukemia, multiple myeloma and acute andchronic leukemias, for example, lymphoblastic, myelogenous, lymphocytic,and myelocytic leukemias.

Specific methods of the invention further comprise the administration ofan additional therapeutic agent (i.e., a therapeutic agent other thanCompound A). Examples of additional therapeutic agents include, but arenot limited to, anti-cancer drugs such as, but are not limited to:alkylating agents, nitrogen mustards, ethylenimines, methylmelamines,alkyl sulfonates, nitrosoureas, triazenes, folic acid analogs,pyrimidine analogs, purine analogs, vinca alkaloids,epipodophyllotoxins, antibiotics, topoisomerase inhibitors andanti-cancer vaccines.

Specific additional therapeutic agents include, but are not limited to:acivicin; aclarubicin; acodazole hydrochloride; acronine; adozelesin;aldesleukin; altretamine; ambomycin; ametantrone acetate;aminoglutethimide; amsacrine; anastrozole; anthramycin; asparaginase;asperlin; azacitidine; azetepa; azotomycin; batimastat; benzodepa;bicalutamide; bisantrene hydrochloride; bisnafide dimesylate; bizelesin;bleomycin sulfate; brequinar sodium; bropirimine; busulfan;cactinomycin; calusterone; caracemide; carbetimer; carboplatin;carmustine; carubicin hydrochloride; carzelesin; cedefingol;chlorambucil; cirolemycin; cisplatin; cladribine; crisnatol mesylate;cyclophosphamide; cytarabine; dacarbazine; dactinomycin; daunorubicinhydrochloride; decitabine; dexormaplatin; dezaguanine; dezaguaninemesylate; diaziquone; docetaxel; doxorubicin; doxorubicin hydrochloride;droloxifene; droloxifene citrate; dromostanolone propionate; duazomycin;edatrexate; eflornithine hydrochloride; elsamitrucin; enloplatin;enpromate; epipropidine; epirubicin hydrochloride; erbulozole;esorubicin hydrochloride; estramustine; estramustine phosphate sodium;etanidazole; etoposide; etoposide phosphate; etoprine; fadrozolehydrochloride; fazarabine; fenretinide; floxuridine; fludarabinephosphate; fluorouracil; flurocitabine; fosquidone; fostriecin sodium;gemcitabine; gemcitabine hydrochloride; hydroxyurea; idarubicinhydrochloride; ifosfamide; ilmofosine; interleukin II (includingrecombinant interleukin II, or rIL2), interferon alfa-2a; interferonalfa-2b; interferon alfa-n1; interferon alfa-n3; interferon beta-I a;interferon gamma-I b; iproplatin; irinotecan hydrochloride; lanreotideacetate; letrozole; leuprolide acetate; liarozole hydrochloride;lometrexol sodium; lomustine; losoxantrone hydrochloride; masoprocol;maytansine; mechlorethamine hydrochloride; megestrol acetate;melengestrol acetate; melphalan; menogaril; mercaptopurine;methotrexate; methotrexate sodium; metoprine; meturedepa; mitindomide;mitocarcin; mitocromin; mitogillin; mitomalcin; mitomycin; mitosper;mitotane; mitoxantrone hydrochloride; mycophenolic acid; nocodazole;nogalamycin; ormaplatin; oxisuran; paclitaxel; pegaspargase; peliomycin;pentamustine; peplomycin sulfate; perfosfamide; pipobroman; piposulfan;piroxantrone hydrochloride; plicamycin; plomestane; porfimer sodium;porfiromycin; prednimustine; procarbazine hydrochloride; puromycin;puromycin hydrochloride; pyrazofurin; riboprine; rogletimide; safingol;safingol hydrochloride; semustine; simtrazene; sparfosate sodium;sparsomycin; spirogermanium hydrochloride; spiromustine; spiroplatin;streptonigrin; streptozocin; sulofenur; talisomycin; tecogalan sodium;tegafur; teloxantrone hydrochloride; temoporfin; teniposide; teroxirone;testolactone; thiamiprine; thioguanine; thiotepa; tiazofurin;tirapazamine; toremifene citrate; trestolone acetate; triciribinephosphate; trimetrexate; trimetrexate glucuronate; triptorelin;tubulozole hydrochloride; uracil mustard; uredepa; vapreotide;verteporfin; vinblastine sulfate; vincristine sulfate; vindesine;vindesine sulfate; vinepidine sulfate; vinglycinate sulfate;vinleurosine sulfate; vinorelbine tartrate; vinrosidine sulfate;vinzolidine sulfate; vorozole; zeniplatin; zinostatin; zorubicinhydrochloride. Other anti-cancer drugs include, but are not limited to:20-epi-1,25 dihydroxyvitamin D3; 5-ethynyluracil; abiraterone;aclarubicin; acylfulvene; adecypenol; adozelesin; aldesleukin; ALL-TKantagonists; altretamine; ambamustine; amidox; amifostine;aminolevulinic acid; amrubicin; amsacrine; anagrelide; anastrozole;andrographolide; angiogenesis inhibitors; antagonist D; antagonist G;antarelix; anti-dorsalizing morphogenetic protein-1; antiandrogen,prostatic carcinoma; antiestrogen; antineoplaston; antisenseoligonucleotides; aphidicolin glycinate; apoptosis gene modulators;apoptosis regulators; apurinic acid; ara-CDP-DL-PTBA; argininedeaminase; asulacrine; atamestane; atrimustine; axinastatin 1;axinastatin 2; axinastatin 3; azasetron; azatoxin; azatyrosine; baccatinIII derivatives; balanol; batimastat; BCR/ABL antagonists;benzochlorins; benzoylstaurosporine; beta lactam derivatives;beta-alethine; betaclamycin B; betulinic acid; bFGF inhibitor;bicalutamide; bisantrene; bisaziridinylspermine; bisnafide; bistrateneA; bizelesin; breflate; bropirimine; budotitane; buthionine sulfoximine;calcipotriol; calphostin C; camptothecin derivatives; canarypox IL-2;capecitabine; carboxamide-amino-triazole; carboxyamidotriazole; CaRestM3; CARN 700; cartilage derived inhibitor; carzelesin; casein kinaseinhibitors (ICOS); castanospermine; cecropin B; cetrorelix; chlorins;chloroquinoxaline sulfonamide; cicaprost; cis-porphyrin; cladribine;clomifene analogues; clotrimazole; collismycin A; collismycin B;combretastatin A4; combretastatin analogue; conagenin; crambescidin 816;crisnatol; cryptophycin 8; cryptophycin A derivatives; curacin A;cyclopentanthraquinones; cycloplatam; cypemycin; cytarabine ocfosfate;cytolytic factor; cytostatin; dacliximab; decitabine; dehydrodidemnin B;deslorelin; dexamethasone; dexifosfamide; dexrazoxane; dexverapamil;diaziquone; didemnin B; didox; diethylnorspermine;dihydro-5-azacytidine; dihydrotaxol, 9-dioxamycin; diphenylspiromustine; docetaxel; docosanol; dolasetron; doxifluridine;droloxifene; dronabinol; duocarmycin SA; ebselen; ecomustine;edelfosine; edrecolomab; eflornithine; elemene; emitefur; epirubicin;epristeride; estramustine analogue; estrogen agonists; estrogenantagonists; etanidazole; etoposide phosphate; exemestane; fadrozole;fazarabine; fenretinide; filgrastim; finasteride; flavopiridol;flezelastine; fluasterone; fludarabine; fluorodaunorunicinhydrochloride; forfenimex; formestane; fostriecin; fotemustine;gadolinium texaphyrin; gallium nitrate; galocitabine; ganirelix;gelatinase inhibitors; gemcitabine; glutathione inhibitors; hepsulfam;heregulin; hexamethylene bisacetamide; hypericin; ibandronic acid;idarubicin; idoxifene; idramantone; ilmofosine; ilomastat;imidazoacridones; imiquimod; immunostimulant peptides; insulin-likegrowth factor-1 receptor inhibitor; interferon agonists; interferons;interleukins; iobenguane; iododoxorubicin; ipomeanol, 4-; iroplact;irsogladine; isobengazole; isohomohalicondrin B; itasetron;jasplakinolide; kahalalide F; lamellarin-N triacetate; lanreotide;leinamycin; lenograstim; lentinan sulfate; leptolstatin; letrozole;leukemia inhibiting factor; leukocyte alpha interferon;leuprolide+estrogen+progesterone; leuprorelin; levamisole; liarozole;linear polyamine analogue; lipophilic disaccharide peptide; lipophilicplatinum compounds; lissoclinamide 7; lobaplatin; lombricine;lometrexol; lonidamine; losoxantrone; lovastatin; loxoribine;lurtotecan; lutetium texaphyrin; lysofylline; lytic peptides;maitansine; mannostatin A; marimastat; masoprocol; maspin; matrilysininhibitors; matrix metalloproteinase inhibitors; menogaril; merbarone;meterelin; methioninase; metoclopramide; MIF inhibitor; mifepristone;miltefosine; mirimostim; mismatched double stranded RNA; mitoguazone;mitolactol; mitomycin analogues; mitonafide; mitotoxin fibroblast growthfactor-saporin; mitoxantrone; mofarotene; molgramostim; monoclonalantibody, human chorionic gonadotrophin; monophosphoryl lipidA+myobacterium cell wall sk; mopidamol; multiple drug resistance geneinhibitor; multiple tumor suppressor 1-based therapy; mustard anticanceragent; mycaperoxide B; mycobacterial cell wall extract; myriaporone;N-acetyldinaline; N-substituted benzamides; nafarelin; nagrestip;naloxone+pentazocine; napavin; naphterpin; nartograstim; nedaplatin;nemorubicin; neridronic acid; neutral endopeptidase; nilutamide;nisamycin; nitric oxide modulators; nitroxide antioxidant; nitrullyn;O6-benzylguanine; octreotide; okicenone; oligonucleotides; onapristone;ondansetron; ondansetron; oracin; oral cytokine inducer; ormaplatin;osaterone; oxaliplatin; oxaunomycin; paclitaxel; paclitaxel analogues;paclitaxel derivatives; palauamine; palmitoylrhizoxin; pamidronic acid;panaxytriol; panomifene; parabactin; pazelliptine; pegaspargase;peldesine; pentosan polysulfate sodium; pentostatin; pentrozole;perflubron; perfosfamide; perillyl alcohol; phenazinomycin;phenylacetate; phosphatase inhibitors; picibanil; pilocarpinehydrochloride; pirarubicin; piritrexim; placetin A; placetin B;plasminogen activator inhibitor; platinum complex; platinum compounds;platinum-triamine complex; porfimer sodium; porfiromycin; prednisone;propyl bis-acridone; prostaglandin J2; proteasome inhibitors; proteinA-based immune modulator; protein kinase C inhibitor; protein kinase Cinhibitors, microalgal; protein tyrosine phosphatase inhibitors; purinenucleoside phosphorylase inhibitors; purpurins; pyrazoloacridine;pyridoxylated hemoglobin polyoxyethylene conjugate; raf antagonists;raltitrexed; ramosetron; ras farnesyl protein transferase inhibitors;ras inhibitors; ras-GAP inhibitor; retelliptine demethylated; rhenium Re186 etidronate; rhizoxin; ribozymes; RII retinamide; rogletimide;rohitukine; romurtide; roquinimex; rubiginone B1; ruboxyl; safingol;saintopin; SarCNU; sarcophytol A; sargramostim; Sdi 1 mimetics;semustine; senescence derived inhibitor 1; sense oligonucleotides;signal transduction inhibitors; signal transduction modulators; singlechain antigen binding protein; sizofiran; sobuzoxane; sodiumborocaptate; sodium phenylacetate; solverol; somatomedin bindingprotein; sonermin; sparfosic acid; spicamycin D; spiromustinc;splenopentin; spongistatin 1; squalamine; stem cell inhibitor; stem-celldivision inhibitors; stipiamide; stromelysin inhibitors; sulfinosine;superactive vasoactive intestinal peptide antagonist; suradista;suramin; swainsonine; synthetic glycosaminoglycans; tallimustine;tamoxifen methiodide; tauromustine; tazarotene; tecogalan sodium;tegafur; tellurapyrylium; telomerase inhibitors; temoporfin;temozolomide; teniposide; tetrachlorodecaoxide; tetrazomine;thaliblastine; thiocoraline; thrombopoietin; thrombopoietin mimetic;thymalfasin; thymopoietin receptor agonist; thymotrinan; thyroidstimulating hormone; tin ethyl etiopurpurin; tirapazamine; titanocenebichloride; topsentin; toremifene; totipotent stem cell factor;translation inhibitors; tretinoin; triacetyluridine; triciribine;trimetrexate; triptorelin; tropisetron; turosteride; tyrosine kinaseinhibitors; tyrphostins; UBC inhibitors; ubenimex; urogenitalsinus-derived growth inhibitory factor; urokinase receptor antagonists;vapreotide; variolin B; vector system, erythrocyte gene therapy;velaresol; veramine; verdins; verteporfin; vinorelbine; vinxaltine;vitaxin; vorozole; zanoterone; zeniplatin; zilascorb; and zinostatinstimalamer.

The invention further encompasses a method of treating or preventingdiseases or disorders ameliorated by the inhibition of PDE4 in a patientwhich comprise administering to a patient in need of such treatment orprevention a therapeutically effective amount of stereomerically pureCompound A, or a pharmaceutically acceptable prodrug, metabolite,polymorph, salt, solvate, hydrate, or clathrate thereof. Disordersameliorated by the inhibition of PDE4 include, but are not limited to,asthma, inflammation, chronic or acute obstructive pulmonary disease,chronic or acute pulmonary inflammatory disease, inflammatory boweldisease, Crohn's Disease, Bechet's Disease, colitis, ulcerative colitisand arthritis or inflammation due to reperfusion. In a preferredembodiment, the disease or disorder to be treated or prevented ischronic obstructive pulmonary disease.

Specific methods of the invention can comprise the administration of anadditional therapeutic agent such as, but not limited to,anti-inflammatory drugs, antihistamines and decongestants. Examples ofsuch additional therapeutic agents include, but are not limited to:antihistamines including, but not limited to, ethanolamines,ethylenediamines, piperazines, and phenothiazines; antiinflammatorydrugs; NSAIDS, including, but not limited to, aspirin, salicylates,acetominophen, indomethacin, sulindac, etodolac, fenamates, tolmetin,ketorolac, diclofenac, ibuprofen, naproxen, fenoprofen, ketoprofen,flurbiprofen, oxaprozin, piroxicam, meloxicam, pyrazolon derivatives;and steriods including, but not limited to, cortical steroids andadrenocortical steroids.

Specific methods of the invention avoid or reduce drug-drug interactionsand other adverse effects associated with agents used in the treatmentof such disorders, including racemic substituted phenylethylsulfones.Without being limited by any theory, stereomerically pure Compound A mayfurther provide an overall improved therapeutic effectiveness, ortherapeutic index, over racemic2-[1-(3-Ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dione.For example, a smaller amount of the drug may in some circumstances beadministered to attain the same level of effectiveness.

As stated above, the active compound of the invention (i.e., Compound A)may be used in the treatment or prevention of a wide range of diseasesand conditions. The magnitude of a prophylactic or therapeutic dose of aparticular active ingredient of the invention in the acute or chronicmanagement of a disease or condition will vary, however, with the natureand severity of the disease or condition, and the route by which theactive ingredient is administered. The dose, and perhaps the dosefrequency, will also vary according to the age, body weight, andresponse of the individual patient. Suitable dosing regimens can bereadily selected by those skilled in the art with due consideration ofsuch factors. In general, the recommended daily dose range for theconditions described herein lie within the range of from about 1 mg toabout 1000 mg per day, given as a single once-a-day dose preferably asdivided doses throughout a day. More specifically, the daily dose isadministered twice daily in equally divided doses. Specifically, a dailydose range should be from about 5 mg to about 500 mg per day, morespecifically, between about 10 mg and about 200 mg per day.Specifically, the daily dose may be administered in 5 mg, 10 mg, 15 mg,20 mg, 25 mg, 50 mg, or 100 mg dosage forms. In managing the patient,the therapy should be initiated at a lower dose, perhaps about 1 mg toabout 25 mg, and increased if necessary up to about 200 mg to about 1000mg per day as either a single dose or divided doses, depending on thepatient's global response. Alternatively, the daily dose is from 0.01mg/kg to 100 mg/kg.

It may be necessary to use dosages of the active ingredient outside theranges disclosed herein in some cases, as will be apparent to those ofordinary skill in the art. Furthermore, it is noted that the clinicianor treating physician will know how and when to interrupt, adjust, orterminate therapy in conjunction with individual patient response.

The phrases “therapeutically effective amount”, “prophylacticallyeffective amount” and “therapeutically or prophylactically effectiveamount,” as used herein encompasses the above described dosage amountsand dose frequency schedules. Different therapeutically effectiveamounts may be applicable for different diseases and conditions, as willbe readily known by those of ordinary skill in the art. Similarly,amounts sufficient to treat or prevent such disorders, but insufficientto cause, or sufficient to reduce, adverse effects associated withracemic2-[1-(3-Ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dioneare also encompassed by the above described dosage amounts and dosefrequency schedules.

4.3. Pharmaceutical Compositions

Pharmaceutical compositions and single unit dosage forms comprisingCompound A, or a pharmaceutically acceptable polymorph, prodrug, salt,solvate, hydrate, or clathrate thereof, are encompassed by theinvention. Individual dosage forms of the invention may be suitable fororal, mucosal (including rectal, nasal, or vaginal), parenteral(including subcutaneous, intramuscular, bolus injection, intraarterial,or intravenous), sublingual, transdermal, buccal, or topicaladministration.

Pharmaceutical compositions and dosage forms of the invention comprisestereomerically pure Compound A, or a pharmaceutically acceptableprodrug, metabolite, polymorph, salt, solvate, hydrate, or clathratethereof. Pharmaceutical compositions and dosage forms of the inventiontypically also comprise one or more pharmaceutically acceptableexcipients.

A particular pharmaceutical composition encompassed by this embodimentcomprises stereomerically pure Compound A, or a pharmaceuticallyacceptable polymorph, prodrug, salt, solvate, hydrate, or clathratethereof, and at least one additional therapeutic agent. Examples ofadditional therapeutic agents include, but are not limited to:anti-cancer drugs and anti-inflammation therapies including, but notlimited to, those listed above in section 4.2.

Single unit dosage forms of the invention are suitable for oral, mucosal(e.g., nasal, sublingual, vaginal, buccal, or rectal), parenteral (e.g.,subcutaneous, intravenous, bolus injection, intramuscular, orintraarterial), or transdermal administration to a patient. Examples ofdosage forms include, but are not limited to: tablets; caplets;capsules, such as soft elastic gelatin capsules; cachets; troches;lozenges; dispersions; suppositories; ointments; cataplasms (poultices);pastes; powders; dressings; creams; plasters; solutions; patches;aerosols (e.g., nasal sprays or inhalers); gels; liquid dosage formssuitable for oral or mucosal administration to a patient, includingsuspensions (e.g., aqueous or non-aqueous liquid suspensions,oil-in-water emulsions, or a water-in-oil liquid emulsions), solutions,and elixirs; liquid dosage forms suitable for parenteral administrationto a patient; and sterile solids (e.g., crystalline or amorphous solids)that can be reconstituted to provide liquid dosage forms suitable forparenteral administration to a patient.

The composition, shape, and type of dosage forms of the invention willtypically vary depending on their use. For example, a dosage form usedin the acute treatment of inflammation or a related disorder may containlarger amounts of one or more of the active ingredients it comprisesthan a dosage form used in the chronic treatment of the same disease.Similarly, a parenteral dosage form may contain smaller amounts of oneor more of the active ingredients it comprises than an oral dosage formused to treat the same disease or disorder. These and other ways inwhich specific dosage forms encompassed by this invention will vary fromone another will be readily apparent to those skilled in the art. See,e.g., Remington's Pharmaceutical Sciences, 18th ed., Mack Publishing,Easton Pa. (1990).

Typical pharmaceutical compositions and dosage forms comprise one ormore excipients. Suitable excipients are well known to those skilled inthe art of pharmacy, and non-limiting examples of suitable excipientsare provided herein. Whether a particular excipient is suitable forincorporation into a pharmaceutical composition or dosage form dependson a variety of factors well known in the art including, but not limitedto, the way in which the dosage form will be administered to a patient.For example, oral dosage forms such as tablets may contain excipientsnot suited for use in parenteral dosage forms. The suitability of aparticular excipient may also depend on the specific active ingredientsin the dosage form.

Lactose-free compositions of the invention can comprise excipients thatare well known in the art and are listed, for example, in the U.S.Pharmacopia (USP) SP (XXI)/NF (XVI). In general, lactose-freecompositions comprise an active ingredient, a binder/filler, and alubricant in pharmaceutically compatible and pharmaceutically acceptableamounts. Preferred lactose-free dosage forms comprise an activeingredient, microcrystalline cellulose, pre-gelatinized starch, andmagnesium stearate.

This invention further encompasses anhydrous pharmaceutical compositionsand dosage forms comprising active ingredients, since water canfacilitate the degradation of some compounds. For example, the additionof water (e.g., 5%) is widely accepted in the pharmaceutical arts as ameans of simulating long-term storage in order to determinecharacteristics such as shelf-life or the stability of formulations overtime. See, e.g., Jens T. Carstensen, Drug Stability: Principles &Practice, 2d. Ed., Marcel Dekker, NY, N.Y., 1995, pp. 379-80. In effect,water and heat accelerate the decomposition of some compounds. Thus, theeffect of water on a formulation can be of great significance sincemoisture and/or humidity are commonly encountered during manufacture,handling, packaging, storage, shipment, and use of formulations.

Anhydrous pharmaceutical compositions and dosage forms of the inventioncan be prepared using anhydrous or low moisture containing ingredientsand low moisture or low humidity conditions. Pharmaceutical compositionsand dosage forms that comprise lactose and at least one activeingredient that comprises a primary or secondary amine are preferablyanhydrous if substantial contact with moisture and/or humidity duringmanufacturing, packaging, and/or storage is expected.

Ah anhydrous pharmaceutical composition should be prepared and storedsuch that its anhydrous nature is maintained. Accordingly, anhydrouscompositions are preferably packaged using materials known to preventexposure to water such that they can be included in suitable formularykits. Examples of suitable packaging include, but are not limited to,hermetically sealed foils, plastics, unit dose containers (e.g., vials),blister packs, and strip packs.

The invention further encompasses pharmaceutical compositions and dosageforms that comprise one or more compounds that reduce the rate by whichan active ingredient will decompose. Such compounds, which are referredto herein as “stabilizers,” include, but are not limited to,antioxidants such as ascorbic acid, pH buffers, or salt buffers.

Like the amounts and types of excipients, the amounts and specific typesof active ingredients in a dosage form may differ depending on factorssuch as, but not limited to, the route by which it is to be administeredto patients. However, typical dosage forms of the invention comprisecompound A, or a pharmaceutically acceptable salt, solvate, clathrate,hydrate, polymorph or prodrug thereof lie within the range of from about1 mg to about 1000 mg per day, given as a single once-a-day dose in themorning but preferably as divided doses throughout the day taken withfood. More specifically, the daily dose is administered twice daily inequally divided doses. Specifically, a daily dose range should be fromabout 5 mg to about 500 mg per day, more specifically, between about 10mg and about 200 mg per day. In managing the patient, the therapy shouldbe initiated at a lower dose, perhaps about 1 mg to about 25 mg, andincreased if necessary up to about 200 mg to about 1000 mg per day aseither a single dose or divided doses, depending on the patient's globalresponse.

4.3.1. Oral Dosage Forms

Pharmaceutical compositions of the invention that are suitable for oraladministration can be presented as discrete dosage forms, such as, butare not limited to, tablets (e.g., chewable tablets), caplets, capsules,and liquids (e.g., flavored syrups). Such dosage forms containpredetermined amounts of active ingredients, and may be prepared bymethods of pharmacy well known to those skilled in the art. Seegenerally, Remington's Pharmaceutical Sciences, 18th ed., MackPublishing, Easton Pa. (1990).

Typical oral dosage forms of the invention are prepared by combining theactive ingredient(s) in an intimate admixture with at least oneexcipient according to conventional pharmaceutical compoundingtechniques. Excipients can take a wide variety of forms depending on theform of preparation desired for administration. For example, excipientssuitable for use in oral liquid or aerosol dosage forms include, but arenot limited to, water, glycols, oils, alcohols, flavoring agents,preservatives, and coloring agents. Examples of excipients suitable foruse in solid oral dosage forms (e.g., powders, tablets, capsules, andcaplets) include, but are not limited to, starches, sugars,micro-crystalline cellulose, diluents, granulating agents, lubricants,binders, and disintegrating agents.

Because of their ease of administration, tablets and capsules representthe most advantageous oral dosage unit forms, in which case solidexcipients are employed. If desired, tablets can be coated by standardaqueous or nonaqueous techniques. Such dosage forms can be prepared byany of the methods of pharmacy. In general, pharmaceutical compositionsand dosage forms are prepared by uniformly and intimately admixing theactive ingredients with liquid carriers, finely divided solid carriers,or both, and then shaping the product into the desired presentation ifnecessary.

For example, a tablet can be prepared by compression or molding.Compressed tablets can be prepared by compressing in a suitable machinethe active ingredients in a free-flowing form such as powder orgranules, optionally mixed with an excipient. Molded tablets can be madeby molding in a suitable machine a mixture of the powdered compoundmoistened with an inert liquid diluent.

Examples of excipients that can be used in oral dosage forms of theinvention include, but are not limited to, binders, fillers,disintegrants, and lubricants. Binders suitable for use inpharmaceutical compositions and dosage forms include, but are notlimited to, corn starch, potato starch, or other starches, gelatin,natural and synthetic gums such as acacia, sodium alginate, alginicacid, other alginates, powdered tragacanth, guar gum, cellulose and itsderivatives (e.g., ethyl cellulose, cellulose acetate, carboxymethylcellulose calcium, sodium carboxymethyl cellulose), polyvinylpyrrolidone, methyl cellulose, pre-gelatinized starch, hydroxypropylmethyl cellulose, (e.g., Nos. 2208, 2906, 2910), microcrystallinecellulose, and mixtures thereof.

Examples of fillers suitable for use in the pharmaceutical compositionsand dosage forms disclosed herein include, but are not limited to, tale,calcium carbonate (e.g., granules or powder), microcrystallinecellulose, powdered cellulose, dextrates, kaolin, mannitol, silicicacid, sorbitol, starch, pre-gelatinized starch, and mixtures thereof.The binder or filler in pharmaceutical compositions of the invention istypically present in from about 50 to about 99 weight percent of thepharmaceutical composition or dosage form.

Suitable forms of microcrystalline cellulose include, but are notlimited to, the materials sold as AVICEL-PH-101, AVICEL-PH-103 AVICELRC-581, AVICEL-PH-105 (available from FMC Corporation, American ViscoseDivision, Avicel Sales, Marcus Hook, Pa.), and mixtures thereof. Anspecific binder is a mixture of microcrystalline cellulose and sodiumcarboxymethyl cellulose sold as AVICEL RC-581. Suitable anhydrous or lowmoisture excipients or additives include AVICEL-PH-103™ and Starch 1500LM.

Disintegrants are used in the compositions of the invention to providetablets that disintegrate when exposed to an aqueous environment.Tablets that contain too much disintegrant may disintegrate in storage,while those that contain too little may not disintegrate at a desiredrate or under the desired conditions. Thus, a sufficient amount ofdisintegrant that is neither too much nor too little to detrimentallyalter the release of the active ingredients should be used to form solidoral dosage forms of the invention. The amount of disintegrant usedvaries based upon the type of formulation, and is readily discernible tothose of ordinary skill in the art. Typical pharmaceutical compositionscomprise from about 0.5 to about 15 weight percent of disintegrant,specifically from about 1 to about 5 weight percent of disintegrant.

Disintegrants that can be used in pharmaceutical compositions and dosageforms of the invention include, but are not limited to, agar-agar,alginic acid, calcium carbonate, microcrystalline cellulose,croscarmellose sodium, crospovidone, polacrilin potassium, sodium starchglycolate, potato or tapioca starch, pre-gelatinized starch, otherstarches, clays, other algins, other celluloses, gums, and mixturesthereof.

Lubricants that can be used in pharmaceutical compositions and dosageforms of the invention include, but are not limited to, calciumstearate, magnesium stearate, mineral oil, light mineral oil, glycerin,sorbitol, mannitol, polyethylene glycol, other glycols, stearic acid,sodium lauryl sulfate, talc, hydrogenated vegetable oil (e.g., peanutoil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil, andsoybean oil), zinc stearate, ethyl oleate, ethyl laureate, agar, andmixtures thereof. Additional lubricants include, for example, a syloidsilica gel (AEROSIL 200, manufactured by W.R. Grace Co. of Baltimore,Md.), a coagulated aerosol of synthetic silica (marketed by Degussa Co.of Plano, Tex.), CAB-O-SIL (a pyrogenic silicon dioxide product sold byCabot Co. of Boston, Mass.), and mixtures thereof. If used at all,lubricants are typically used in an amount of less than about 1 weightpercent of the pharmaceutical compositions or dosage forms into whichthey are incorporated.

4.3.2. Delayed Release Dosage Forms

Active ingredients of the invention can be administered by controlledrelease means or by delivery devices that are well known to those ofordinary skill in the art. Examples include, but are not limited to,those described in U.S. Pat. Nos. 3,845,770; 3,916,899; 3,536,809;3,598,123; and 4,008,719, 5,674,533, 5,059,595, 5,591,767, 5,120,548,5,073,543, 5,639,476, 5,354,556, and 5,733,566, each of which isincorporated herein by reference. Such dosage forms can be used toprovide slow or controlled-release of one or more active ingredientsusing, for example, hydropropylmethyl cellulose, other polymer matrices,gels, permeable membranes, osmotic systems, multilayer coatings,microparticles, liposomes, microspheres, or a combination thereof toprovide the desired release profile in varying proportions. Suitablecontrolled-release formulations known to those of ordinary skill in theart, including those described herein, can be readily selected for usewith the active ingredients of the invention. The invention thusencompasses single unit dosage forms suitable for oral administrationsuch as, but not limited to, tablets, capsules, gelcaps, and capletsthat are adapted for controlled-release.

All controlled-release pharmaceutical products have a common goal ofimproving drug therapy over that achieved by their non-controlledcounterparts. Ideally, the use of an optimally designedcontrolled-release preparation in medical treatment is characterized bya minimum of drug substance being employed to cure or control thecondition in a minimum amount of time. Advantages of controlled-releaseformulations include extended activity of the drug, reduced dosagefrequency, and increased patient compliance. In addition,controlled-release formulations can be used to affect the time of onsetof action or other characteristics, such as blood levels of the drug,and can thus affect the occurrence of side (e.g., adverse) effects.

Most controlled-release formulations are designed to initially releasean amount of drug (active ingredient) that promptly produces the desiredtherapeutic effect, and gradually and continually release of otheramounts of drug to maintain this level of therapeutic or prophylacticeffect over an extended period of time. In order to maintain thisconstant level of drug in the body, the drug must be released from thedosage form at a rate that will replace the amount of drug beingmetabolized and excreted from the body. Controlled-release of an activeingredient can be stimulated by various conditions including, but notlimited to, pH, temperature, enzymes, water, or other physiologicalconditions or compounds.

4.3.3. Parenteral Dosage Forms

Parenteral dosage forms can be administered to patients by variousroutes including, but not limited to, subcutaneous, intravenous(including bolus injection), intramuscular, and intraarterial. Becausetheir administration typically bypasses patients' natural defensesagainst contaminants, parenteral dosage forms are preferably sterile orcapable of being sterilized prior to administration to a patient.Examples of parenteral dosage forms include, but are not limited to,solutions ready for injection, dry products ready to be dissolved orsuspended in a pharmaceutically acceptable vehicle for injection,suspensions ready for injection, and emulsions.

Suitable vehicles that can be used to provide parenteral dosage forms ofthe invention are well known to those skilled in the art. Examplesinclude, but are not limited to: Water for Injection USP; aqueousvehicles such as, but not limited to, Sodium Chloride Injection,Ringer's Injection, Dextrose Injection, Dextrose and Sodium ChlorideInjection, and Lactated Ringer's Injection; water-miscible vehicles suchas, but not limited to, ethyl alcohol, polyethylene glycol, andpolypropylene glycol; and non-aqueous vehicles such as, but not limitedto, corn oil, cottonseed oil, peanut oil, sesame oil, ethyl oleate,isopropyl myristate, and benzyl benzoate.

Compounds that increase the solubility of one or more of the activeingredients disclosed herein can also be incorporated into theparenteral dosage forms of the invention.

4.3.4. Transdermal, Topical, and Mucosal Dosage Forms

Transdermal, topical, and mucosal dosage forms of the invention include,but are not limited to, ophthalmic solutions, sprays, aerosols, creams,lotions, ointments, gels, solutions, emulsions, suspensions, or otherforms known to one of skill in the art. See, e.g., Remington'sPharmaceutical Sciences, 16th and 18th eds., Mack Publishing, Easton Pa.(1980 & 1990); and Introduction to Pharmaceutical Dosage Forms, 4th ed.,Lea & Febiger, Philadelphia (1985). Dosage forms suitable for treatingmucosal tissues within the oral cavity can be formulated as mouthwashesor as oral gels. Further, transdermal dosage forms include “reservoirtype” or “matrix type” patches, which can be applied to the skin andworn for a specific period of time to permit the penetration of adesired amount of active ingredients.

Suitable excipients (e.g., carriers and diluents) and other materialsthat can be used to provide transdermal, topical, and mucosal dosageforms encompassed by this invention are well known to those skilled inthe pharmaceutical arts, and depend on the particular tissue to which agiven pharmaceutical composition or dosage form will be applied. Withthat fact in mind, typical excipients include, but are not limited to,water, acetone, ethanol, ethylene glycol, propylene glycol,butane-1,3-diol, isopropyl myristate, isopropyl palmitate, mineral oil,and mixtures thereof to form lotions, tinctures, creams, emulsions, gelsor ointments, which are non-toxic and pharmaceutically acceptable.Moisturizers or humectants can also be added to pharmaceuticalcompositions and dosage forms if desired. Examples of such additionalingredients are well known in the art. See, e.g., Remington'sPharmaceutical Sciences, 16th and 18th eds., Mack Publishing, Easton Pa.(1980 & 1990).

Depending on the specific tissue to be treated, additional componentsmay be used prior to, in conjunction with, or subsequent to treatmentwith active ingredients of the invention. For example, penetrationenhancers can be used to assist in delivering the active ingredients tothe tissue. Suitable penetration enhancers include, but are not limitedto: acetone; various alcohols such as ethanol, oleyl, andtetrahydrofuryl; alkyl sulfoxides such as dimethyl sulfoxide; dimethylacetamide; dimethyl formamide; polyethylene glycol; pyrrolidones such aspolyvinylpyrrolidone; Kollidon grades (Povidone, Polyvidone); urea; andvarious water-soluble or insoluble sugar esters such as Tween 80(polysorbate 80) and Span 60 (sorbitan monostearate).

The pH of a pharmaceutical composition or dosage form, or of the tissueto which the pharmaceutical composition or dosage form is applied, mayalso be adjusted to improve delivery of one or more active ingredients.Similarly, the polarity of a solvent carrier, its ionic strength, ortonicity can be adjusted to improve delivery. Compounds such asstearates can also be added to pharmaceutical compositions or dosageforms to advantageously alter the hydrophilicity or lipophilicity of oneor more active ingredients so as to improve delivery. In this regard,stearates can serve as a lipid vehicle for the formulation, as anemulsifying agent or surfactant, and as a delivery-enhancing orpenetration-enhancing agent. Different salts, hydrates or solvates ofthe active ingredients can be used to further adjust the properties ofthe resulting composition.

4.3.5. Kits

Typically, active ingredients of the invention are preferably notadministered to a patient at the same time or by the same route ofadministration. This invention therefore encompasses kits which, whenused by the medical practitioner, can simplify the administration ofappropriate amounts of active ingredients to a patient.

A typical kit of the invention comprises a unit dosage form of compoundA, or a pharmaceutically acceptable salt, solvate, hydrate, clathrate,polymorph or prodrug thereof, and a unit dosage form of a second activeingredient. Examples of second active ingredients include, but are notlimited to, those listed in section 4.2 above.

Kits of the invention can further comprise devices that are used toadminister the active ingredient(s). Examples of such devices include,but are not limited to, syringes, drip bags, patches, and inhalers.

Kits of the invention can further comprise pharmaceutically acceptablevehicles that can be used to administer one or more active ingredients.For example, if an active ingredient is provided in a solid form thatmust be reconstituted for parenteral administration, the kit cancomprise a sealed container of a suitable vehicle in which the activeingredient can be dissolved to form a particulate-free sterile solutionthat is suitable for parenteral administration. Examples ofpharmaceutically acceptable vehicles include, but are not limited to:Water for Injection USP; aqueous vehicles such as, but not limited to,Sodium Chloride Injection, Ringer's Injection, Dextrose Injection,Dextrose and Sodium Chloride Injection, and Lactated Ringer's Injection;water-miscible vehicles such as, but not limited to, ethyl alcohol,polyethylene glycol, and polypropylene glycol; and non-aqueous vehiclessuch as, but not limited to, corn oil, cottonseed oil, peanut oil,sesame oil, ethyl oleate, isopropyl myristate, and benzyl benzoate.

5. EXAMPLES 5.1. Example 1 SYNTHESIS OF2-[1-(3-ETHOXY-4-METHOXYPHENYL)-2-METHYLSULFONYLETHYL]-4-ACETYLAMINOISOINDOLINE-13-DIONE

A stirred solution of1-(3-ethoxy-4-methoxyphenyl)-2-methylsulfonylethylamine (1.0 g, 3.7mmol) and 3-acetamidophthalic anhydride (751 mg, 3.66 mmol) in aceticacid (20 mL) was heated at reflux for 15 h. The solvent was removed invacuo to yield an oil. Chromatography of the resulting oil yielded theproduct as a yellow solid (1.0 g, 59% yield): mp, 144° C.; ¹H NMR(CDCl₃) δ1.47 (t, J=7.0 Hz, 3H, CH₃), 2.26 (s, 3H, CH₃), 2.88 (s, 3H,CH₃), 3.75 (dd, J=4.4, 14.3 Hz, 1H, CHH), 3.85 (s, 3H, CH₃), 4.11 (q,J=7 Hz, 2H, CH2), 5.87 (dd, J=4.3, 10.5 Hz, 1H, NCH), 6.82-6.86 (m, 1H,Ar), 7.09-7.11 (m, 2H, Ar), 7.47 (d, J=7 Hz, 1H, Ar), 7.64 (t, J=8 Hz,1H, Ar), 8.74 (d, J=8 Hz, 1H, Ar), 9.49 (br s, 1H, NH); ¹³C NMR (CDCl₃)δ14.61, 24.85, 41.54, 48.44, 54.34, 55.85, 64.43, 111.37, 112.34,115.04, 118.11, 120.21, 124.85, 129.17, 130.96, 136.01, 137.52, 148.54,149.65, 167.38, 169.09, 169.40; Anal Calc'd. for C₂₂H₂₄NO₇S: C, 57.38;H, 5.25; N, 6.08. Found: C, 57.31; H, 5.34; N, 5.83.

5.2. Example 2 SYNTHESIS OF(+)2-[1-(3-ETHOXY-4-METHOXYPHENYL)-2-METHYLSULFONYLETHYL]-4ACETYLAMINOISOINDOLINE-1,3-DIONE

Preparation of 3-aminopthalic acid

10% Pd/C (2.5 g), 3-nitrophthalic acid (75.0 g, 355 mmol) and ethanol(1.5 L) were charged to a 2.5 L Parr hydrogenator, under a nitrogenatmosphere. Hydrogen was charged to the reaction vessel for up to 55psi. The mixture was shaken for 13 hours, maintaining hydrogen pressurebetween 50 and 55 psi. Hydrogen was released and the mixture was purgedwith nitrogen 3 times. The suspension was filtered through a celite bedand rinsed with methanol. The filtrate was concentrated in vacuo. Theresulting solid was reslurried in ether and isolated by vacuumfiltration. The solid was dried in vacuo to a constant weight, affording54 g (84% yield) of 3-aminopthalic acid as a yellow product. ¹H-NMR(DMSO-d6) δ: 3.17 (s, 2H), 6.67 (d, 1H), 6.82 (d, 1H), 7.17 (t, 1H),8-10 (brs, 2H). ¹³C-NMR (DMSO-d6) δ: 112.00, 115.32, 118.20, 131.28,135.86, 148.82, 169.15, 170.09.

Preparation of 3-acetamidophthalic anhydride

A 1 L 3-necked round bottom flask was equipped with a mechanicalstirrer, thermometer, and condenser and charged with 3-aminophthalicacid (108 g, 596 mmol) and acetic anhydride (550 mL). The reactionmixture was heated to reflux for 3 hours and cooled to ambienttemperature and further to 0-5° C. for another 1 hour. The crystallinesolid was collected by vacuum filtration and washed with ether. Thesolid product was dried in vacuo at ambient temperature to a constantweight, giving 75 g (61% yield) of 3-acetamidopthalic anhydride as awhite product. ¹H-NMR (CDCl₃) δ: 2.21 (s, 3H), 7.76 (d, 1H), 7.94 (t,1H), 8.42 (d, 1H), 9.84 (s, 1H).

Resolution of2-(3-ethoxy-4-methoxyphenyl)-1-(methylsulphonyl)-eth-2-ylamine

A 3 L 3-necked round bottom flask was equipped with a mechanicalstirrer, thermometer, and condenser and charged with2-(3-ethoxy-4-methoxyphenyl)-1-(methylsulphonyl)-eth-2-ylamine (137.0 g,500 mmol), N-acetyl-L-leucine (52 g, 300 mmol), and methanol (1.0 L).The stirred slurry was heated to reflux for 1 hour. The stirred mixturewas allowed to cool to ambient temperature and stirring was continuedfor another 3 hours at ambient temperature. The slurry was filtered andwashed with methanol (250 mL). The solid was air-dried and then dried invacuo at ambient temperature to a constant weight, giving 109.5 g (98%yield) of the crude product (85.8% ee). The crude solid (55.0 g) andmethanol (440 mL) were brought to reflux for 1 hour, cooled to roomtemperature and stirred for an additional 3 hours at ambienttemperature. The slurry was filtered and the filter cake was washed withmethanol (200 mL). The solid was air-dried and then dried in vacuo at30° C. to a constant weight, yielding 49.6 g (90% recovery) of(S)-2-(3-ethoxy-4-methoxyphenyl)-1-(methylsulphonyl)-eth-2-ylamine-N-acetyl-L-leucinesalt (98.4% ee). Chiral HPLC ( 1/99 EtOH/20 mM KH₂PO₄@pH 7.0, UltronChiral ES-OVS from Agilent Technologies, 150 mm×4.6 mm, 0.5 mL/min.,@240 mm): 18.4 min (S-isomer, 99.2%), 25.5 min (R-isomer, 0.8%).

Preparation of Compound A

A 500 mL 3-necked round bottom flask was equipped with a mechanicalstirrer, thermometer, and condenser. The reaction vessel was chargedwith (S)-2-(3-ethoxy-4-methoxyphenyl)-1-(methylsulphonyl)-eth-2-yl amineN-acetyl-L-leucine salt (25 g, 56 mmol, 98% ee), 3-acetamidophthalicanhydride (12.1 g 58.8 mmol), and glacial acetic acid (250 mL). Themixture was refluxed over night and then cooled to <50° C. The solventwas removed in vacuo, and the residue was dissolved in ethyl acetate.The resulting solution was washed with water (250 mL×2), saturatedaqueous NaHCO₃ (250 mL×2), brine (250 mL×2), and dried over sodiumsulphate. The solvent was evaporated in vacuo, and the residuerecrystallized from a binary solvent containing ethanol (150 mL) andacetone (75 mL). The solid was isolated by vacuum filtration and washedwith ethanol (100 mL×2). The product was dried in vacuo at 60° C. to aconstant weight, affording 19.4 g (75% yield) ofS-{2-[1-(3-ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-aminoisoindoline-1,3-dionewith 98% ee. Chiral HPLC ( 15/85 EtOH/20 mM KH₂PO₄ @pH 3.5, UltronChiral ES-OVS from Agilent Technology, 150 mm×4.6 mm, 0.4 mL/min., @240nm): 25.4 min (S-isomer, 98.7%), 29.5 min (R-isomer, 1.2%). ¹H-NMR(CDCl₃) δ: 1.47 (t, 3H), 2.26 (s, 3H), 2.87 (s, 3H), 3.68-3.75 (dd, 1H),3.85 (s, 3H), 4.07-4.15 (q, 2H), 4.51-4.61 (dd, 1H), 5.84-5.90 (dd, 1H),6.82-8.77 (m, 6H), 9.46 (s, 1H). ¹³C-NMR (DMSO-d6) δ: 14.66, 24.92,41.61, 48.53, 54.46, 55.91, 64.51, 11.44, 112.40, 115.10, 118.20,120.28, 124.94, 129.22, 131.02, 136.09, 137.60, 148.62, 149.74, 167.46,169.14, 169.48.

5.3. Example 3 TNF-α Inhibition

Human Whole Blood LPS-Induced TNF-α Assay

The ability of compounds to inhibit LPS-induced TNF-α production byhuman whole blood was measured essentially as described below for theLPS-induced TNF-α assay in human PBMC, except that freshly drawn wholeblood was used instead of PBMC. (George Muller, et al. 1999, Bioorganic& Medicinal Chemistry Letters 9; 1625-1630.) Human whole bloodLPS-induced TNF-α IC₅₀-294 nM

Mouse LPS-Induced Serum TNF-α Inhibition

Compounds were tested in this animal model according to previouslydescribed methods (Corral et al. 1996, Mol. Med. 2:506-515). MouseLPS-induced serum TNF-α inhibition (ED₅₀, mg/kg, p.o.)=0.05.

LPS-Induced TNF-α Production

Lipopolysaccharide (LPS) is an endotoxin produced by gram-negativebacteria such as E. coli which induces production of manypro-inflammatory cytokines, including TNF-α. In peripheral bloodmononuclear cells (PBMC), the TNF-α produced in response to LPS isderived from monocytes, which comprise approximately 5-20% of the totalPBMC. Compounds were tested for the ability to inhibit LPS-induced TNF-αproduction from human PBMC as previously described (Muller et al. 1996,J. Med. Chem. 39:3238). PBMC from normal donors were obtained by FicollHypaque (Pharmacia, Piscataway, N.J., USA) density centrifugation. Cellswere cultured in RPMI (Life Technologies, Grand Island, N.Y., USA)supplemented with 10% AB± human serum (Gemini Bio-products, Woodland,Calif., USA), 2 mM L-glutamine, 100 U/ml penicillin, and 100 μg/mlstreptomycin (Life Technologies).

PBMC (2×10⁵ cells) were plated in 96-well flat-bottom Costar tissueculture plates (Corning, N.Y., USA) in triplicate. Cells were stimulatedwith LPS (Sigma, St. Louis, Mo., USA) at 100 ng/ml in the absence orpresence of compounds. Compounds (Celgene Corp., Warren, N.J., USA) weredissolved in DMSO (Sigma) and further dilutions were done in culturemedium immediately before use. The final DMSO concentration in allsamples was 0.25%. Compounds were added to cells 1 hour before LPSstimulation. Cells were incubated for 18-20 hours at 37° C. in 5% CO₂and supernatants were then collected, diluted with culture medium andassayed for TNF-α levels by ELISA (Endogen, Boston, Mass., USA).LPS-induced TNF-α IC₅₀=77 nM.

IL-1β-Induced TNF-α Production

During the course of inflammatory diseases, TNF-α production is oftenstimulated by the cytokine IL-1β, rather than by bacterially derivedLPS. Compounds were tested for the ability to inhibit IL-1β-inducedTNF-α production from human PBMC as described above for LPS-inducedTNF-α production, except that the PBMC were isolated from sourceleukocyte units (Sera-Tec Biologicals, North Brunswick, N.J., USA) bycentrifugation on Ficoll-Paque Plus (Amersham Pharmacia, Piscataway,N.J., USA), plated in 96-well tissue culture plates at 3×10⁵ cells/wellin RPMI-1640 medium (BioWhittaker, Walkersville, Md., USA) containing10% heat-inactivated fetal bovine serum (Hyclone), 2 mM L-glutamine, 100U/ml penicillin, and 100 mg/ml streptomycin (complete medium),pretreated with compounds at 10, 2, 0.4, 0.08, 0.016, 0.0032, 0.00064,and 0 μM in duplicate at a final DMSO concentration of 0.1% at 37° C. ina humidified incubator at 5% CO₂ for 1 hour, then stimulated with 50ng/ml recombinant human IL-1β (Endogen) for 18 hours. IL-β-induced TNF-αIC₅₀=83 nM.

5.4. Example 4 PDE Selectivity

PDE1, 2, 3, 5, and 6 Enzyme Assays

The specificity of compounds for PDE4 was assessed by testing at asingle concentration (10 μM) against bovine PDE1, human PDE2, PDE3, andPDE5 from human platelets (Hidaka and Asano 1976, Biochem. Biophys. Acta429:485, and Nicholsen et al. 1991, Trends Pharmaco. Sci. 12:19), andPDE6 from bovine retinal rod outer segments (Baehr et al. 1979, J. Biol.Chem. 254:11669, and Gillespie et al. 1989, Mol. Pharm. 36:773). Resultsare listed in Table 1.

PDE7 Enzyme Assay

PDE7 is a cAMP-selective PDE expressed mainly in T cells and in skeletalmuscle. T cell-derived cytokines such as IL-2 and IFN-γ are potentiallyregulatable via PDE7 inhibition. PDE7 was purified from Hut78 human Tcells by anion exchange chromatography as previously described (Bloomand Beavo 1996, Proc. Natl. Acad. Sci. USA 93:14188-14192). Compoundswere tested against the PDE7 preparation in the presence of 10 nM cAMPas described for PDE4 in Table 1 below. TABLE 1 Racemic CompoundCompound Compound A B* PDE Inhibition PDE4 IC₅₀ (from U937 81.8 73.5 611cells) (nM) PDE1 (% inhib at 10 μM)  9% 23% 27% PDE2 (% inhib at 10 μM)19%  6% 10% PDE3 (% inhib at 10 μM) 21% 20% 31% PDE5 (% inhib at 10 μM) 3%  3% −9% PDE6 (% inhib at 10 μM) ND −6% 10% PDE7 IC₅₀ (nM) 2211020500 ND PDE Specificity Ratios from above data (*fold)PDE4/PDE1 >2700 >500 >50 PDE4/PDE2 >800 >10000 >260PDE4/PDE3 >670 >1200 >45 PDE4/PDE5 >12000 >30000 >39000 PDE4/PDE6ND >40000 >250 PDE7 IC₅₀/PDE4 IC₅₀ 270 279 ND*Compound B is the opposite enantiomer of Compound A.

5.5. Example 5 PDE4 Inhibition

PDE4 (U937 Cell-Derived) Enzyme Assay

PDE4 enzyme was purified from U937 human monocytic cells by gelfiltration chromatography as previously described (Muller et al. 1998,Bioorg. & Med Chem Lett 8:2669-2674). Phosphodiesterase reactions werecarried out in 50 mM Tris HCl pH 7.5, 5 mM MgCl₂, 1 μM cAMP, 10 nM[³H]-cAMP for 30 min at 30° C., terminated by boiling, treated with 1mg/ml snake venom, and separated using AG-1XS ion exchange resin(BioRad) as described (Muller et al. 1998, Bioorg. & Med Chem Lett8:2669-2674). Reactions consumed less than 15% of available substrate.Results are listed in Table 1.

5.6. Example 6 Human T Cell Assays

SEB-Induced IL-2 and IFN-γ Production

Staphylococcal Enterotoxin B (SEB) is a superantigen derived fromgram-positive bacteria Staphylococcus aureus. SEB provides a convenientphysiological stimulus specific for T cells expressing particular T cellreceptor Vβ chains. Human PBMC (consisting of approximately 50% T cells)were isolated from source leukocyte units as described above and platedin 96-well tissue culture plates at 3×10⁵ cells/well in complete medium,pretreated with compounds at 10, 2, 0.4, 0.08, 0.016, 0.0032, 0.00064,and 0 μM in duplicate at a final DMSO concentration of 0.1% at 37° C. ina humidified incubator at 5% CO₂ for 1 hour, then stimulated with 100ng/ml SEB (Sigma Chemical Co., St. Louis, Mo., USA) for 18 hours. IL-2and IFN-γ levels were measured by ELISA (R&D Systems, Minneapolis,Minn., USA). IL-2IC₅₀=291 nM. IFN-γ IC₅₀=46 nM.

5.7. Example 6 cAMP Elevation Assays

PGE₂-Induced cAMP Elevation

Prostaglandin E₂ (PGE₂) binds to prostanoid receptors on monocytes, Tcells and other leukocytes and consequently elevates intracellular cAMPlevels, resulting in inhibition of cellular responses. The combinationof PGE₂ and a PDE4 inhibitor synergistically elevates cAMP levels inthese cell types, and the elevation of cAMP in PBMC caused by PDE4inhibitors in the presence of PGE₂ is proportional to the inhibitoryactivity of that PDE4 inhibitor. Intracellular cAMP was measured inhuman PBMC as follows. PBMC were isolated as described above and platedin 96-well plates at 1×10⁶ cells per well in RPMI-1640. The cells werepre-treated with compounds at 100, 10, 1, 0.1, 0.01, and 0 μM in a finalconcentration of 2% DMSO in duplicate at 37° C. in a humidifiedincubator at 5% CO₂ for one hour. The cells were then stimulated withPGE₂ (10 μM) (Sigma) for 1 h. The cells were lysed with HCl, 0.1N finalconcentration to inhibit phosphodiesterase activity and the plates werefrozen at −20° C. The cAMP produced was measured using cAMP (low pH)Immunoassay kit (R&D Systems). PBMC cAMP EC₅₀ for racemate is 3.09 μCM.PBMC cAMP EC₅₀ for Compound A is 1.58 μM.

Elevation of cAMP in human neutrophils was measured as follows. PBMCwere removed from source leukocytes (Sera-Tec Biologicals) bycentrifugation on Ficoll-Paque Plus (Amersham Pharmacia). The resultingerythrocyte/polymorphonuclear cell (PMN) pellet was resuspended inHank's Balanced Salt Solution (BioWhittaker) and mixed with an equalvolume of 3% Dextran T-500 (Amersham Pharmacia) in 0.9% saline.Erythrocytes were allowed to sediment for 20 minutes, and the PMN wereremoved and centrifuged at 120 rpm for 8 minutes at 4° C. The remainingerythrocytes were lysed in cold 0.2% saline for 30 seconds, and thecells restored to isotonicity by the addition of an equal volume of 1.6%saline. The PMN were centrifuged at 1200 rpm for 8 minutes at 4° C.,then resuspended in RPMI-1640 and assayed for cAMP elevation asdescribed for PBMC above. PMN were found to be approximately 74%CD18/CD11b⁺, 71% CD16⁺CD9′ neutrophils by flow cytometzy on aFACSCalibur (Becton Dickinson, San Jose, Calif., USA). Results are shownin Table 2.

fMLF-Induced LTB4 Production

N-formyl-methionine-leucine-phenylalanine (fMLF) is a bacteriallyderived peptide that activates neutrophils to rapidly degranulate,migrate, adhere to endothelial cells, and release leukotriene LTB4, aproduct of arachidonic acid metabolism and itself a neutrophilchemoattractant. Compounds were tested for the ability to blockfMLF-induced neutrophil LTB4 production as previously described(Hatzelmann and Schudt 2001, J. Pharm. Exp. Ther. 297:267-279), with thefollowing modifications. Neutrophils were isolated as described aboveand resuspended in phosphate-buffered saline without calcium ormagnesium (BioWhittaker) containing 10 mM HEPES pH7.2 and plated in96-well tissue culture plates at a concentration of 1.7×10⁶ cells/well.Cells were treated with 50 μM thimerosal (Sigma)/1 mM CaCl₂/1 mM MgCl₂for 15 minutes at 37° C. 5% CO₂, then treated with compounds at 1000,200, 40, 8, 1.6, 0.32, 0.064, and 0 nM in a final DMSO concentration of0.01% in duplicate for 10 minutes. Neutrophils were stimulated with 1 μMfMLF for 30 minutes, then lysed by the addition of methanol (20% finalconcentration) and frozen in a dry ice/isopropanol bath for 10 minutes.lysates were stored at −70° C. until the LTB4 content was measured bycompetitive LTB4 ELISA (R&D Systems). Results are shown in Table 2.

Zymosan-Induced IL-8 Production

Zymosan A, or the heat-killed yeast Saccharomyces cerevisiae, binds tothe adhesion molecule Mac-1 on the neutrophil surface and triggersphagocytosis, cell activation and IL-8 production. Zymosan-induced IL-8production was measured as previously described (Au et al. 1998, Brit.J. Pharm. 123:1260-1266) with the following modifications. Humanneutrophils were purified as described above, plated in 96-well tissueculture plates at 3×10⁵ cells/well in complete medium, treated withcompounds at 10, 2, 0.4, 0.08, 0.016, 0.0032, 0.00064, and 0 μM induplicate in a final DMSO concentration of 0.1% for 1 hour at 37° C. 5%CO₂. Neutrophils were then stimulated with unopsonized, boiled Zymosan A(Sigma) at 2.5×10⁵ particles/well for 18 hours. Supernatants wereharvested and tested for IL-8 by ELISA (R&D Systems). Results are shownin Table 2.

fMLF-Induced CDI8/CD11 Expression

CD18/CD11b (Mac-1) expression on neutrophils was measured as previouslydescribed (Derian et al. 1995, J. Immunol.:154:308-317) with thefollowing modifications. Neutrophils were isolated as described above,then resuspended in complete medium at 1×10⁶ cells/ml, pretreated withcompounds at 10, 1, 0.1, 0.01, and 0 μM in duplicate at a final DMSOconcentration of 0.1% for 10 minutes at 37° C. 5% CO₂. Cells were thenstimulated with 30 nM fMLF for 30 minutes and then chilled to 4° C.Cells were treated with rabbit IgG (Jackson ImmunoResearch Labs, WestGrove, Pa., USA) (10 μg/1×10⁶ cells) to block Fc receptors, stained withCD18-FITC and CD11b-PE (Becton Dickinson), and analyzed by flowcytometry on a FACSCalibur. CD18/CD11b expression (mean fluorescence) inthe absence of stimulation was subtracted from all samples to obtaininhibition curves and calculate IC₅₀s. Results are shown in Table 2.

fMLF-Induced Adhesion to HUVEC

Human umbilical vein endothelial cells (HUVEC) were used as a substratefor neutrophil adhesion as previously described (Derian et al. 1995, J.Immunol.: 154:308-317) with the following modifications. HUVEC cellswere obtained from Anthrogenesis (Cedar Knolls, N.J., USA), andneutrophils were not treated with cytochalasin B. Cells were treatedwith compounds at 10, 1, 0.1, 0.01, 0.001, and 0 μM in a final DMSOconcentration of 0.1% in duplicate for 10 minutes, stimulated with 500μM fMLF for 30 minutes, and washed twice with PBS before measuringfluorescence on an FLX800 plate reader (Bio-Tek Instruments, Winooski,Vt., USA). Results are shown in Table 2. TABLE 2 Human Neutrophil AssaysRacemic (all values in nM) Compound Compound A PGE₂-induced cAMP EC₅₀12589 4570 fMLF-induced LTB4 IC₅₀ 20.1 2.48 Zymosan-induced IL-8 IC₅₀ ND94 fMLF-induced CD18 expression IC₅₀ ND 390 fMLF-induced CD11bexpression IC₅₀ ND 74 fMLF-induced adhesion to HUVEC IC₅₀ ND 150

5.8. Example 8 Aqueous Solubility

Equilibrium solubilities were measured in pH 7.4 aqueous buffer. The pH7.4 buffer was prepared by adjusting the pH of a 0.07 M NaH₂PO₄ solutionto 7.4 with 10 N NaOH. The ionic strength of the solution was 0.15. Atleast 1 mg of powder was combined with 1 ml of buffer to make >1 mg/mlmixture. These samples were shaken for >2 hours and left to standovernight at room temperature. The samples were then filtered through a0.45-μm Nylon syringe filter that was first saturated with the sample.The filtrate was sampled twice, consecutively. The filtrate was assayedby HPLC against standards prepared in 50% methanol. Compound A has3.5-fold greater aqueous solubility than the racemic mixture. Measuredsolubility Compound A=0.012 mg/mL; racemic mixture=0.0034 mg/mL.

5.9. Example 8 LPS-Induced Lung Neutrophilia Ferret Model

The conscious ferret model has been used to investigateanti-inflammatory, emetic and behavioral effects of PDE4 inhibitors whenadministered by the oral (p.o.) route. From these experiments, atherapeutic index (TI) for each PDE4 inhibitor may be determined. The TIhas been calculated by dividing the threshold dose for causing emeticepisodes and behavioral changes by the anti-inflammatory dose (dose thatcauses 50% inhibition of the LPS-induced neutrophilia).

Animal Husbandry

Male ferrets (Mustela Pulorius Euro, weighing 1-2 kg). Ferrets weresupplied either by Bury Green Farm or Misay Consultancy. Followingtransport, the animals were allowed to acclimatize in the holding roomsfor a period of not less than 7 days. The Diet comprised SDS diet Cpelleted food given ad Jib with Whiskers cat food given 3 times perweek. Water was pasteurized animal grade drinking water and was changeddaily.

Dosing with PDE4 Inhibitor

PDE4 inhibitors were administered orally (p.o.), at doses initially of1-10 mg/kg, but subsequently up to 30 mg/kg in order to establishwhether the TI was 10 or higher, and/or at lower doses to establish theminimum dose to cause 50% inhibition of neutrophilia. Ferrets werefasted overnight but allowed free access to water. The animals wereorally dosed with vehicle or PDE4 inhibitor using a 15 cm dosing needlethat was passed down the back of the throat into the esophagus. Afterdosing, the animals were returned to holding cages fitted with Perspexdoors to allow observation, and given free access to water. Afterdosing, the animals were constantly observed and any emesis orbehavioural changes were recorded. The animals were allowed access tofood 60-90 minutes after p.o. dosing

Exposure to LPS

Thirty minutes after p.o. dosing with compound or vehicle control, theferrets were placed into sealed Perspex containers and exposed to anaerosol of LPS (100 μg/ml) for 10 minutes. Aerosols of LPS weregenerated by a nebulizer (DeVilbiss, USA) and this was directed into thePerspex exposure chamber. Following a 10 minute exposure period, theanimals were returned to the holding cages and allowed free access towater, and at a later stage, food. Observation continued for a period ofat least 2.5 hours post p.o. dosing and emetic episodes and behavioralchanges were recorded.

Bronchoalveolar Ravage

Six hours after LPS exposure the animals were killed by overdose ofsodium pentobarbitone administered intraperitoneally. The trachea wasthen cannulated with polypropylene tubing and the lungs lavaged twicewith 20 ml heparinized (10 units/ml) phosphate buffered saline (PBS).

Blood Sampling/Tissue Removal

A terminal blood sample (10 ml) was removed by trans-thoracic cardiacpuncture. The blood was spun at 2500 rpm for 15 minutes and the plasmaremoved and stored at −20° C. The brain also removed and frozen at −20°C. for analysis of compound content.

Cell Counts

The bronchoalveolar lavage (BAL) samples were centrifuged at 1500 rpmfor 5 minutes. The supernatant was removed and the resulting cell pelletre-suspended in 1 ml PBS. A cell smear of the re-suspended fluid wasprepared and stained with Leishmans stain to allow differential cellcounting. A total cell count was made using the remaining re-suspendedsample. From this, the total number of neutrophils in the BAL wasdetermined.

Parameters Measured:

1. % Inhibition of LPS-induced pulmonary neutrophilia.

2. Emetic episodes—the number of vomits and retches were counted.

3. Behavioral changes—the following behavioral effects were noted:salivation, panting, mouth clawing, flattened posture, ataxia, archedback and backward walking. Any behavioral changes were semi-quantifiedby applying a severity rating (mild, moderate or severe).

4. The TI was calculated as the highest dose found to not cause emeticepisodes divided by the lowest dose found to inhibit pulmonaryneutrophilia by 50% or more.

The effect of Compound A on LPS-induced neutrophilia in the lungs ofconscious ferrets is demonstrated in FIG. 1.

Emesis and Behavioral Changes

Following p.o. dosing of the PDE4, the ferrets were observed for atleast 2 hours and emetic episodes (vomits and retches) and behavioralchanges were recorded.

No emetic episodes (retching or vomiting) were observed in the ferretspre-treated p.o. with the relevant vehicle (acetone/cremophor/distilledwater). In a small proportion of the control-treated animals ( 7/22),mild behavioral changes (lip licking and backward walking) were seen.

Compound A (0.1-3 mg/kg, p.o.), caused no emetic episodes (retching andvomiting). Some behavioral changes (flattened posture, lip licking andbackward walking) were observed and classified as mild. At 10 mg/kg in2/6 ferrets, some retching but no frank emesis was observed along withsalivation and behavioral changes (scored as mild or moderate). At thehighest dose tested (30 mg/kg) moderate to marked emesis was observed in¾ animals along with pronounced behavioral changes. These data aresummarized in Table III. TABLE III Conscious ferret: Emetic episodes andbehavioural changes following oral administration of Compound A.Treatment/dose Mouth Flattened Backward (mg/kg) Vomits RetchesSalivation Panting clawing posture Ataxia Lip licking walking VehicleNone None None None None None None Mild Mild (acetone/cremophor/ (6/22)(7/22) dist.H2O) Compound A None None None None None Mild None Mild Mild(0.1 mg/kg) (2/5) (4/5) (3/5) Compound A None None None None None MildNone Mild Mild (0.3 mg/kg) (2/6) (3/6) (4/6) Compound A None None NoneNone None Mild None Mild Mild (1.0 mg/kg) (2/6) (6/6) (4/6) Compound ANone None None None Mild Marked None Mild Moderate (3.0 mg/kg) (1/8)(7/8) (2/8) (5/8) Compound A None Mild Mild None Mild Marked NoneModerate Marked (10 mg/kg) (2/6) (1/6) (1/6) (6/6) (5/6) (6/6) CompoundA Moderate Marked Moderate Mild Marked Marked Mild Moderate Mild (30mg/kg) (3/4) (3/4) (3/4) (1/4) (4/4) (4/4) (3/4) (4/4) (2/4)

Animals were observed for up to 3 hours following dosing. Numbers inparentheses refer to the number of animals that responded The numbers ofanimals in each group range from 4-22.

Therapeutic Index Calculation

From these experiments, a therapeutic index (TI) was determined for eachcompound by dividing the threshold dose for inducing emetic episodes bythe ED₅₀ value for inhibiting the pulmonary neutrophilia. The TIcalculation is summarized in Table IV. Compound A had a TI of 12,causing no emetic episodes at an anti-inflammatory dose of 1 mg/kg.TABLE IV Summary of the effective doses (ED₅₀) for inhibition ofLPS-induced pulmonary neutrophilia and induction of emesis and thetherapeutic index derived from these values. Inhibition of LPS-inducedneutrophilia Threshold emetic Therapeutic Compound (ED₅₀ mg/kg) dose(mg/kg) index Compound A 0.8 10 12

5.10. Example 9 200 MG Dosage Capsule

Table V illustrates a batch formulation and single dosage formulationfor a 200 mg Compound A single dose unit, i.e., about 40 percent byweight, in a size #0 capsule. TABLE V Formulation for 200 mg capsulePercent Quantity Quantity Material By Weight (mg/tablet) (kg/batch)Compound A 40.0%   200 mg 16.80 kg Pregelatinized Corn 9.5% 297.5 mg24.99 kg Starch, NF5 Magnesium Stearate 0.5%  2.5 mg  0.21 kg Total100.0%   500 mg 42.00 kg

The pregelatinized corn starch (SPRESS B-820) and Compound A componentsare passed through a 710 μm screen and then are loaded into a DiffusionMixer with a baffle insert and blended for 15 minutes. The magnesiumstearate is passed through a 210 μm screen and is added to the DiffusionMixer. The blend is then encapsulated in a size #0 capsule, 500 mg percapsule (8400 capsule batch size) using a Dosator type capsule fillingmachine.

5.11. Example 10 100 MG Oral Dosage Form

Table VI illustrates a batch formulation and a single dose unitformulation containing 100 mg of Compound A. TABLE VI Formulation for100 mg tablet Percent Quantity Material by Weight Quantity (mg/tablet)(kg/batch) Compound A  40% 100.00 20.00 Microcrystalline 53.5%  133.7526.75 Cellulose, NF Pluronic F-68 4.0% 10.00 2.00 SurfactantCroscarmellose 2.0% 5.00 1.00 Sodium Type A, NF Magnesium 0.5% 1.25 0.25Stearate, NF Total 100.0%  250.00 mg 50.00 kg

The microcrystalline cellulose, croscarmellose sodium, and Compound Acomponents are passed through a #30 mesh screen (about 430V to about655μ). The Pluronic F-68® (manufactured by JRH Biosciences, Inc. ofLenexa, Kans.) surfactant is passed through a #20 mesh screen (about457μ to about 1041μ). The Pluronic F-68® surfactant and 0.5 kgs ofcroscarmellose sodium are loaded into a 16 qt. twin shell tumble blenderand are mixed for about 5 minutes. The mix is then transferred to a 3cubic foot twin shell tumble blender where the microcrystallinecellulose is added and blended for about 5 minutes. The thalidomide isadded and blended for an additional 25 minutes. This pre-blend is passedthrough a roller compactor with a hammer mill attached at the dischargeof the roller compactor and moved back to the tumble blender. Theremaining croscarmellose sodium and magnesium stearate is added to thetumble blender and blended for about 3 minutes. The final mixture iscompressed on a rotary tablet press with 250 mg per tablet (200,000tablet batch size).

5.12. Example 11 Aerosol Dosage Form

A concentrate is prepared by combining Compound A, and a 12.6 kg portionof the trichloromonofluormethane in a sealed stainless steel vesselequipped with a high shear mixer. Mixing is carried out for about 20minutes. The bulk suspension is then prepared in the sealed vessel bycombining the concentrate with the balance of the propellants in a bulkproduct tank that is temperature controlled to 21° to 27° C. andpressure controlled to 2.8 to 4.0 BAR. 17 ml aerosol containers whichhave a metered valve which is designed to provide 100 inhalations of thecomposition of the invention. Each container is provided with thefollowing: Compound A 0.0120 g trichloromonofluoromethane 1.6939 gdichlorodifluoromethane 3.7175 g dichlorotetrafluoroethane 1.5766 gtotal 7.0000 g

While the invention has been described with respect to the particularembodiments, it will be apparent to those skilled in the art thatvarious changes and modifications may be made without departing from thespirit and scope of the invention as defined in the claims. Suchmodifications are also intended to fall within the scope of the appendedclaims.

1-55. (canceled)
 56. A method of treating erythema nodosum leprosum inleprosy, rhematoid spondylitis, hepatitis or congestive heart failurewhich comprises administering to a patient a therapeutically effectiveamount of stereomerically pure(+)-2-[1-(3-Ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dione,or a pharmaceutically acceptable prodrug, polymorph, salt, or solvatethereof.
 57. The method of claim 56 further comprising administering toa patient in need of such treatment a therapeutically effective amountof an antihistamine, anti-inflammatory drug, non-steroidanti-inflammatory drug or steroid.
 58. The method of claim 56 or 57wherein the patient is a mammal.
 59. The method of claim 56 or 57wherein the stereomerically pure(+)-2-[1-(3-Ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dioneis administered parenterally, transdermally, mucosally, nasally,buccally, sublingually, or orally.
 60. The method of claim 59 whereinthe stereomerically pure(+)-2-[1-(3-Ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dioneis administered orally.
 61. The method of claim 60 wherein thestereomerically pure(+)-2-[1-(3-Ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dioneis administered orally in a tablet or capsule form.
 62. The method ofclaim 61 wherein the therapeutically effective amount is from about 1 mgto about 1000 mg per day.
 63. The method of claim 62 wherein thetherapeutically effective amount is from about 5 mg to about 500 mg perday.
 64. The method of claim 63 wherein the therapeutically effectiveamount is from about 10 mg to about 200 mg per day.
 65. The method ofclaim 63 wherein the therapeutically effective amount is from about 20mg per day.
 66. The method of claim 63 wherein the therapeuticallyeffective amount is from about 20 mg twice daily.